Parliamentary reforms in the nineteenth and early twentieth centuries Essay Example for Free

Parliamentary reforms in the nineteenth and early twentieth centuries Essay How far would you agree that fear of popular hostility was the main reason why governments enacted parliamentary reforms in the nineteenth and early twentieth centuries? The reform movement first started to achieve mass support in the 1760s; during the years of the French Revolution it even appealed to the members of the anti-reform Whig party but as war with France started, the authorities feared revolution as had happened in France and so used repressive measures in a bid to stamp out these new ideas. It was in 1815 however that the reform movement began to become more significant when widespread unemployment attracted the masses to the idea of reform. But as conditions improved during the 1820s the pressure for reform decreased as Cobbett said, I defy you to agitate a man on a full stomach. The reform movement was not unified nor was it on a national scale. There were different groups urging for changes the most extensive of these were the radicals, such men as Robert Paine. They campaigned for universal male suffrage, annual parliaments, equal electoral districts, a secret ballot, the payment of MPs and the abolition of property qualifications for MPs. There were some more radical that others however and along with little agreement on what should be the best policy of reform they had little success on parliament. There were also calls for reform from more moderate radicals and members of the Whigs that called for measures for more limited reform such as disfranchising the worst of the rotten boroughs and giving representation to larger towns. The lack of reform prior to the 1832 Act can be explained by the lack of unity of the reformers and their less that significant support in the country but more importantly the Tories anti-reform majority in parliament and the small number of pro-reform Whigs and radicals. The economic slump of 1829 and an increase in poor harvests caused unemployment and distress for many working class families and thus made them more susceptible to reformist ideology. This period saw the reform movement revived by such radicals as Cobbett and Hunt; the BPU, a reform organisation was also founded by Thomas Attwood which provided a pressure group aligning the lower and middle class people this provided the opportunity for others to be created throughout the country during 1829-30 with the middle and working classes increasingly cooperating together; the combination of the two provided a class alliance that the Tories were fearful of. The death of the pro-Tory King George IV necessitated a general election in which the Tories majority was heavily reduced and the continuation of Wellingtons government became uncertain with the increase of Whig seats. This was due to the pro-reform attitude of the country especially within the counties and open boroughs. The election saw the return of Henry Brougham in a seat for Yorkshire; he was more popular in the country than any Whig leader and had promised to initiate parliamentary reform. With the reform movements continued support, other agitation began to emerge in the country. The Swing Riots of Southern England involved burning hayricks and breaking machinery which they blamed for reducing employment for farm workers. Although easily suppressed, the Whigs, worried of unrest and encouraged by the Tory decline, announced their intention to introduce reform legislation in the Commons; Wellington however continually expressed his belief that the existing system possessed the full and entire confidence of the country. In November 1830 the defeat of the Tory government in the Commons brought an end to their 20 years of rule. This opened the door to the Whigs who had already expressed intention in reforming parliament who formed a minority government. The Whigs aim of producing this legislation was a measure large enough to satisfy public opinion but to also provide resistance to further innovation and to uphold the authority of the Aristocracy and the existing Whig government. Although they wanted to remove the most blatant abuses, they were heavily concerned with preserving as much as possible the social and political status quo. Their strategy was to remedy the grievances of the middle classes thereby gaining their support and dividing the middle-working class alliance of the reform movement which posed huge problems for the government of left unresolved. The Bill made no concessions to the radicals and working classes meaning that post-reform agitation would almost certainly continue. During the Bills progression through parliament, agitation continued. Political unions organised demonstrations, riots occurred in Nottingham and Bristol and further violence seemed possible. This extra-parliamentary agitation only strengthened the Whig governments determination in passing the Bill. When King William IV refused to create more Whig peers to carry the Bill through the Lords, Grey resigned and Wellington took office again. This resulted in the Days of May where nationwide protests and demonstrations made some fearful of revolution; reformers also threatened an economic crisis by withdrawing gold from the banks to stop the Duke, go for gold. With Wellingtons failure at forming a government, and the huge public opinion in favour of reform, the King had no choice but to create these necessary peers the Tory majority in the House of Lords however conceded and the Bill was passed. It can therefore be seen that although reform was intended by the Whigs, there was still fear of an uprising that could have swayed members votes, especially in the Commons. Further parliamentary reform was inevitable the Whigs had recognised the huge public opinion in favour of reform and were now in a majority in the House of Commons. Lord Althorp even warned Grey in 1833 that without popular measures, the Reform Act will lead to revolution. It is therefore clear that the fear of uprisings was still apparent to many even after the Act was passed. Although the next Reform Act wasnt passed until 1867, there was still popular agitation in the country. This especially came from the Chartist movement whose demands would have essentially made Britain into a democracy; their demands were however ignored by parliament largely based on the fact that anti-reformer Palmerston was in charge for much of the period between the Acts. With an increasing number of reform legislation being passed both social and economic the country was becoming more democratic and with this came factors in determining parliamentary reform. As well as radical demands for reform continuing, a new factor of party political opportunism was also present it was in the Conservative parties own interests to pass the Second Reform Act. As had happened prior to the 1832 Act, 1866 saw economic problems which increased social discontent and fuelled the calls for reform this is evident in the huge surge of membership to the Reform Union and Reform League. In 1866 both organisations one middle class, the other working started working together, creating the same threat that the Tories had feared back in 1830. It was the Liberal party that introduced the first Bill to parliament, with them seeing the opportunity in enfranchising selective members of working class who were already in favour of the party. The radicals backed the Bill but believed that the measure was too limited; many right-wing Liberals however thought the opposite that the Bill would enfranchise too many of the working class. The Conservatives saw the opportunity of the split ideology and the Liberal party and worked with the right-wingers in defeating the Bill in June of 1866; the Liberal government resigned, giving way to a minority Conservative administration. Despite his own parties opposition to reform, Disraeli introduced his own reform Bill based purely on self-interests in gaining the support of the proposed boroughs to be enfranchised. Moreover, Disraeli also claimed that he intended to destroy the present agitation in the country. As well as party political interests, agitation was still ongoing with radicals and organisations pushing for reform. A peaceful demonstration by the Reform League in London had erupted in violence in July 1966; this, along with further riots, convinced many MPs on the need for reform. Disraelis Bill introduced in March 1867 was more moderate than the Liberal one before it, with the intention that those who opposed the last Bill will accept this one. Rather than it being defeated, Disraeli was willing to make the Bill even more radical than the one prior it was their certainty that redistributing seats would return a Conservative majority that made members within the party support the Bill, along with the radical MPs. It can therefore be seen that self-interests of parties was the main factor in bringing about the Second Reform Act rather than agitation in the country, which was the case during the 1830s. Calls for a secret ballot had been apparent since the late eighteenth century, it was even considered during the Reform Bill in 1831. It wasnt however agitation that led to the Ballot Act of 1872, it was the election of 1868. Violence, corruption and intimidation had occurred as in every election prior but what was surprising was the publicity the election received with journalists commenting on the extent to which violence was a factor. The government set up a committee in 1870 to look into this with their report favouring a system of secret voting. Although the Conservatives apposed such a measure, they saw the opportunity in getting radical support so pushed ahead with the Bill. Intimidation still continued even with an increased electorate and secret voting it simply meant that candidates had to spend more to win over the voters. Secret voting did however mean that voters would not necessarily vote for the candidate they had received money from; this, along with radical calls for reform, urged governments of the time to act. The 1883 Corrupt and Illegal Practices Act passed with both Conservative and Liberal support it limited the expenditure of elections (fell by three-quarters in the 1885 election) and in turn reduced corruption and violence. It was parties interests saving them the huge expense of elections that again prevailed over agitation to pass reform. After the 1867 Act, most politicians had accepted that further parliamentary reform was inevitable, even if they didnt support it. It was therefore important to ensure that reform was in the parties interests, as had occurred in 1867. It was again party self-interests that determined the Third Reform Act Gladstone agreed to support moderate reform believing he would gain popularity in the country. A Liberal Bill was introduced in 1884 with the Conservative majority of the House of Lords, led by Salisbury, believing that although an extension of the county franchise would be beneficial to the Liberals, a redistribution of seats would prevent them from gaining a sufficient majority. By this time conflict had reappeared again, not regarding reform but the abolition of the House of Lords. The riots that followed were not on the same level as in 1832 and the government had little to worry about. There was no significant extra-parliamentary pressure for reform, it was the parties acting in self-interest for themselves in trying to pass the Bill that really counted. A compromise was finally reached the Arlington Street compact in which both parties got what they wanted the Liberals an extension of the franchise and the Conservatives the redistribution of seats. With each successive Act making the House of Commons more democratic, the House of Lords remained essentially aristocratic, with many radicals proposing the upper House be reformed. There were repeated calls for the House to be abolished, especially during the Liberals Home Rule Bill, which was thrown out by the Lords. The election of 1906 produced a Liberal majority of over 200 seats, with the Lords still a Conservative stronghold poised to block key Liberal measures in the following years. The Parliament Bill was introduced in 1910, but the Lords were reluctant to pass such a measure that would removed their power of veto. Liberal Prime Minister Asquith therefore urged the King to create more peers to secure a Liberal majority insisting that there was a serious state of unrest among the middle classes that could lead to harming the crown. The Bill was reintroduced to Parliament with the House of Lords finally accepting the proposals in August 1911 rather then see the House swamped by Liberal peers. The 1911 Parliament Act can therefore be seen as very little to do with agitation in the country and more of the fact of the increasing hostility between the Liberal government and the Conservation Lords. Although the electoral system was now more representative of the people than ever before, there were still huge numbers of people who were denied the vote women for example made up half of the population but still had no proper political voice. The system was therefore still criticised, with the Liberal party calling for further reform including manhood suffrage and the abolition of plural voting. It was from the womens suffrage movement that significant pressure on parliament after 1885 emerged. Parliament found it increasingly difficult to ignore calls for reform from organisations such as the National Union of Womens Suffrage and the Womens Social and Political Union, both of which held demonstrations and were increasing in popularity. They began more militant activities including vandalism and attacks on MPs which only helped to alienate potential supporters, it was in fact the more moderate groups that found the greater success. Once again it came down to party politics with the Liberal party in 1905 believing that enfranchising women on the same basis as men would create a majority middle-class, who were more than likely to vote Conservative. The reform movement virtually disappeared during the outbreak of the war, which reinforces the claim that the Fourth Reform Act passed in 1918 was a result of the parties, and not extra-parliamentary pressure. The Liberals and Labour both supported universal male suffrage by now, with the Conservatives apposing the idea at first but hoping such enfranchised men would vote Conservative. Women were also partly included in the act not because of suffragette pressure, but because before the war a majority of MPs favoured extending the vote to them as long as it was in their parties interests. It can therefore be seen that virtually no agitation or pressure was put on parliament during the time leading up to the 1918 Act, it was simply due to the self-interests of the parties with each benefiting from a particular aspect of the Bill.

Puc18 Plasmid Engineering

Puc18 Plasmid Engineering Abstract The objective of the experiment was to engineer a pUC18 plasmid so that it contained a kanomycin resistance gene in its multiple cloning site and to transform it into cells. The kanomycin resistance gene was obtained from a pKAN plasmid. The desired plasmid was constructed by digesting pUC18 and pKAN with the same restriction enzymes,(BamHI and HindIII) and religating the products to give the engineered pUC18. The created plasmid was then transformed into E.coli strains DH5ÃŽ ±. The strains that contained the engineered plasmid were selected using two methods of selection. According to the indirect method of selection the percentage of competent cells transformed with the plasmids was 0.063% which is a low number. According to the direct method of selection on the other hand no cells were transformed. In conclusion even though some colonies with the engineered plasmids were obtained the percentage of cells transformed was very low. Also, the indirect method of selection gives better results for selection of desired strains. Introduction Bacteria can carry antibiotic resistance genes either in their chromosomes or extrachromosomally in phage or a plasmid(Hausner and de Jong 2010). B-galactosidase is an enzyme involved into the cleavage of lactose into glucose and galactose and is encoded by the lac Z gene of the lac operon.(Glick et al 2010) The lac operon is prevented from being transcribed through repression of the lac promoter. Activation of this promoter can be done by the addition of lactose or isopropyl-ÃŽ ²-D-thiogalactopyranoside(IPTG) to the medium. Lactose and IPTG simply prevent binding of the lac repressor(the product of the Lac I gene) to the promoter. (Glick et al 2010) In the following experiment plasmids pUC18 and pKAN are used to provide the genes to be transformed into bacteria. pUC18 is 2686 base pairs(bp) long and contains a bacterial origin of replication, an ampicillin resistance gene, a lacI gene, a segment of the lac Z gene encoding part of B-galactosidase(which breaks down X-gal) and a multiple cloning sequence(MCS) that is within the lac Z gene.(Glick et al 2010) The lac Z gene encoded by the plasmid is part of the B-galactosidase protein which complements a gene carried by the Escheria. coli chromosomally thus forming a functional B-galactosidase.(Glick et al 2010) If a DNA segment is cloned in the MCS then the lacZ gene will be interrupted and will not give rise to a functional protein. If that occurs then the Bacteria transformed with the plasmid will not break down5-bromo-4-chloro-3-indolyl-ÃŽ ²-D-ÃŽ ²-galactosidase( X-gal) present in the plates. When X-gal is broken down by ÃŽ ²-galactosidase it turns blue whereas when it is not bro ken down it stays white. This color differentiation is a way to tell if there has been any DNA incorporated in the MCS of pUC18. Finally in order for the ÃŽ ²-galactosidase in pUC18 to be transcribed, IPTG has to be present in the medium so that the lac operon can be induced.(Glick et al 2010) pKAN plasmids can serve as sources for the kanomycin resistance gene. In the following experiment the kanaomycin resistance gene will be inserted in the MCS of pUC18. pKAN contains an origin of replication, a kanomycin resistance gene and multiple restriction sites.(Hausner and de Jong 2010) More importantly it contains only one BamHI and HindIII recognition sites in the whole plasmid which flank the kanomycin resistance gene.(Hausner and de Jong) This allows researchers to cut out the antibiotic resistance gene by simply using BamHI and HindIII producing only two fragments of DNA: the gene and the rest of the plasmid. Once experimenters have inserted the pKAN gene into the MCS of pUC18 and transformed the E.coli strains they need a way to select for the desired plasmid. There are two methods to select for the desired those colonies: the direct method and the indirect method. The direct selection method involves spread plating transformed strains into plates containing both the antibiotic ampicillin and kanomycin. (Hausner and de Jong 2010) Since the pUC18 plasmid confers amplicillin resistance(Glick et al 2010) and the kan gene confers kanomycin resistance (Hausner and de Jong 2010) then only the cells that contain Puc18 with the kanomycin resistance gene should be able to grow in these plates. The indirect method on the other hand is a two step selection process. In the first step the transformed strains are plated onto LB plates containing ampicillin and X-gal.(Hausner and de Jong 2010) Only the cells that have up-taken pUC18 will grow since they will be resistant to ampicillin. Furthermore cell s that contain pUC18 with inserted DNA in the MCS will produce white colonies since they cant produce a functional ÃŽ ²-galactosidase. Cells that give rise to blue colonies will have up-taken pUC18 without any DNA inserted in their MCS since they are able to break down X-Gal. (Glick et al 2010) To select the cells with pUC18 containing the kanomycin resistance gene the white colonies are plated in plates containing kanomycin. Only the cells that have the kanomycin resistance gene in their pUC18 will grow.(Hausner and de Jong 2010) The objectives of the following experiment include the construction of a pUC18 plasmid containing the kanomycin resistance gene in the MCS, the transformation of that plasmid into the E.coli DH5ÃŽ ± cells and the selection of the cells containing the engineered plasmid. If both pUC18 and pKAN plasmids are digested with BamHI and HindIII and the digests are ligated then a plasmid which contains both kanomycin and ampicillin resistance genes should be produced; consequently cells transformed with the engineered plasmid should be resistant to both antibiotics. Materials and Methods Plasmid extraction and plasmid engineering pUC18 and pKAN plasmids were extracted from the DH5ÃŽ ± and MM294 E.coli strains respectively using a DNA isolation kit as described by (Hausner and de Jong 2010). Confirmation for proper extraction was done through agarose gel electrophoresis by running the extracted DNA in a 0.7% gel at 100V for 1 hour. The gene containing kanomycin resistance from pKAN was cloned into pUC18. The restriction digests to do the cloning were prepared as described in Table 2 in (Hausner and de Jong 2010). After plasmid digestion the kanomycin resistance gene was inserted into the multiple cloning sequence of pUC18 in a ligation reaction using the enzyme ligase and the reaction was allowed to go to completion for 24 hours at room temperature. The ligation reactions were set up according to table 3 in (Hausner and de Jong 2010) E.coli transformation and strain selection E.coli strain DH5ÃŽ ± was sub-cultured for 1 hour at 37Â °C. The cells were then made competent by washing them in 10mM CaCl. Next cells were transformed with three different combinations of plasmids. The set of cells in tube 1 was transformed with uncut pUC18 DNA. The set of cells in tube 2 was transformed with cut pUC18. Cells in tube 3 were transformed with pUC18 containing the cloned pKAN resistance and finally cells in tube 4 were transformed with just water as a negative control. The transformation procedure has been described in (Hausner and de Jong 2010). Transformed cells from all tubes were spread plated onto LB+carb+X-gal plates for indirect selection. Furthermore cells from tube 3 were plated onto LB+carb+ kan plates for direct selection of cells containing pUC18 with the insert from pKAN. To determine the density of competent cells cells dilutions of , and were prepared. The two highest dilutions were plated onto LB plates. All the plates were incubated at 37Â °C and they were allowed to grow for ~24 hours. After the colonies had grown on plates plate they were counted and their numbers were recorded. White and blue colonies from the LB+carb+X-gal plates were then streaked onto LB + kan plates to obtain the colonies that had the kanomycin resistance gene incorporated in the MCS. For more information on the procedure refer to Experiments in Biotechnology Laboratory Manual (Hausner and de Jong 2010) Results Extraction of plasmids from E.coli strains Figure 1 contains the image of the 0.7% agarose gel in which the isolated plasmids Puc18 and pKAN were run to check for product. As it can be seen in lane 1 a lot of Puc18 was extracted from the DH5ÃŽ ± strain. Less plasmid DNA was collected for pKAN from the MM294 strain since the band in lane 2 is of much weaker intensity. There is more than one band in lane two. The additional bands represent additional plasmids isolated from the bacteria. Calculation of Competent cell density Table 1 shows the dilutions performed on the competent cells in order to calculate their cell density. It also shows the number of colonies on the plates that were spread plated with dilution 2 and dilution 3. The results for the dilution were not used for cell density calculation since less than 30 colonies grew on the plate. Dilution was used to calculate the cell density because the number of colonies was between 30 and 300. Indirect method of selection Cells plated from tubes 2 and 3 were used to calculate the % of transformed cells. Every colony represents a single transformed cell since it can be assumed the every colony has arisen from a single cell. Furthermore for tube 3 since five plates were spread plated the percentage of the transformed cells was obtained by using the average amount of colonies for all five plates. Calculation the percentage of transformed cells in tube 2: %of transformed cells= x 100 =0.0045% of cells transformed Calculation of transformed cells in tube 3 Average for blue colonies: = 58.6 ≈ 59 blue colonies Average for white colonies = 11.4 ≈ 11colonies Total number of colonies = 59 blue colonies + 11 blue colonies = 70 colonies in total Both blue and white colonies from tube 3 represent transformed cells since they both up-took plasmid DNA whether it was just pUC18 or pUC18+kanomycin resistance gene. Therefore since every colony came from a single cell there were 70 cells in total that were transformed from 100Â µl of media spread plated in each plate. % of transformed cells in tube 3: %of transformed cells= x 100 =0.063% of cells transformed Direct selection of clones containing the kanomycin gene: No colonies grew on LB + carb + kan plates. That means that there were no cells that were transformed with the engineered plasmid. Furthermore an accurate number for % of transformed cell could not have been calculated even if cells had grown in these plates. That is because this selection method takes into account only the cells that were trasformend with pUC18 which contained the kanomycin resistance gene and not the cells that were transformed with only pUC18. Discussion Isolation of plasmids from cells The optimal results for the gel would have been to see one strong band at ~2.7 kb representing pUC18 and one strong band at 4.2 kb which represents pKAN. For the pKAN lane there is more than one band seen. Those bands represent different sized plasmids that were also isolated from the cell. Since there was no DNA ladder on the gel it cannot be concluded what plasmid the lanes represent but the only thing that can be concluded is that there was plasmid DNA isolated from both the DH5ÃŽ ± and the MM294 strains which most likely was pUC18 and pKAN. In order to conclude whether pUC18 and pKAN plasmids were isolated from the bacteria the students should be provided next time with a DNA ladder in order to determine the sizes of the lanes. Indirect selection method The cells from tube 1 were transformed with un-digested pUC18. The cells from this tube represented a positive control for transformation. The colonies in the plates were all blue and they were too many to count. The reason for the high number of colonies was that these cells were transformed with undigested plasmids which are all stable and all allow bacteria to carry information extrachromosomally, making the transformation percentage of competent cells very high. All the cells from tube 1 produce blue colonies. That is because they all had a functional B-galactisidase since no genes were cloned into the multiple cloning site located within the lacZ gene. The cells from tube 2 were transformed with digested pUC18 plasmid. The cells from this tube represented a negative control for kanomycin resistance gene cloning. Tube 2 gave rise to very few colonies in comparison to tube 1 because the cells in tube 2 were transformed with unstable DNA. pUC18 had been previously digested with HinDIII and BamHI and a lot of plasmid did not re-ligate and for that reason the DNA was unstable. Since the DNA was unstable it was not able to maintain the ampicillin resistance gene in bacteria and consequently the strains were not able to grow in carbonicillin plates. As a result the number of percent transformed cells was as low as 0.0045%. The cells from tube 4 were transformed with sterile water i.e no DNA. These cells represented the negative control for transformation. Because no DNA was inserted in them none of the cells contained the ampicillin resistance gene and as expected none grew in the plates containing carbomicillin. The cells from tube 3 were transformed using pUC18 that contained insertion on the MCS as well as pUC18 that didnt. All five plates that were spread plated with E.coli from tube 3 contained blue colonies as well as white ones. The reason for the color difference is that the blue colonies contained a functional ÃŽ ²-galactosidase whereas the white ones didnt. The functional ÃŽ ²-galactosidase in the blue colonies was due to the fact that no DNA was inserted in the MCS to interrupt the lacZ gene. The white colonies on the other hand did not contain a functional ÃŽ ²-galactosidase since they had a DNA insertion in their multiple cloning site, which interrupted the lacZ gene. Consequently they could not break down X-gal. However just because they had a DNA insertion in their MCS it did not mean that they contained the kanomycin resistance gene. They might have contained the rest of the pKAN plasmid. As a result the white colonies needed to be streaked into plates that selected for kanomy cin resistance. If the cells then grew on LB + Kan plates and they also originated from white colonies on LB + Carb + X-gal plates then they contained a Puc19 plasmid with a kanomycin resistance gene inserted in the MCS. The percentage of transformed cells was also not very high: 0.063%. A way to improve this would be to maybe increase the molarity of the CaCl solution to make the cells more competent. Direct selection method According to the direct method of selection there were no cells that were transformed. This is contradictory to the results obtained from the indirect method of selection. This error could have been produced because of either improper spread plating of plates or because of improper transformation procedure. Also the conditions in the LB + carb + kan plates could have been too harsh(two antibiotics) for the bacteria to pick up growth even if they were resistant to both antibiotics. In following experiments it is better to use the indirect selection method since it seems more successful in selecting desired strains. Comparison of direct VS indirect selection methods The direct and indirect selection methods have both advantages as well as disadvantages. The main disadvantage of indirect selection is that it takes longer since it contains two steps and each step takes at least a day for completion. The main advantage is that if done correctly, the indirect selection methods gives very accurate selection for the desired cells. The reason for that is that first it selects for colonies that just have an insertion in the MCS and this tells the researcher that some type of cloning has occurred in plasmids. The second step then selects for the colonies that contain pUC18 with the kanomycin resistance gene inserted in the MCS. Thus the criterion of indirect selection is that cells have both pUC18 with an inserted DNA in MCS and also have kanomycin resistance. The colonies that grow in the second step fulfill both the criteria. The main advantage of the direct method is that it takes a shorter time to complete and it also uses up less equipment which can also save researchers some money. The main disadvantage with this selection is that it has a higher chance of giving false positives. Direct selection does not select for strains that have DNA inserted in the MCS of Puc18 but only selects for strains that have ampicillin and kanomycin resistance. Therefore the strains that grow in LB + carb + kan plates might have both pUC18 and pKAN plasmids but not the kanomycin resistance gene inserted in the pUC18 MCS. Those strains would still be able to grow since they still have both ampicillin and kanomycin resistance. However the genes would on different plasmids and not on the engineered one. Therefore even though the indirect selection method is longer it is more accurate in selecting the desired strains for this experiment. In conclusion, according to the indirect selection the desired plasmid was engineered by digesting both pUC18 and pKAN with HindIII and BamHI. Also when selecting for cells transformed with pUC18 it is better to employ the indirect method of selection because it gives more accurate results. Question 1: Although both lanes contain plasmid DNA, why doesnt the DNA appear to be in the same location in both lanes? The DNA does not appear in the same location in both lanes because pUC18 and pKAN are of different sizes. pUC18 is 2686 base pais long whereas pKAN is 4194 base pairs long.(Hausner and de Jong 2010) Because pUC18 is of smaller size it will travel farther from the wells than pKAN. Question 2: How would you verify that the transformed cells actually contain the carb/kan plasmid that was used for transformation? One accurate way would be to isolate the plasmid DNA from the transformad cells and run it on an agarose gel. If the kanomycin resistance gene was inserted into pUC18 then on the gel one will be able to see a band of the size 4548 base pairs which is different from both the pUC18 and the pKAN plasmids. The size of the created plasmid was calculated the following way by obtaining the information from (Hausner and de Jong 2010): To find the size of kanomycin resistance gene inserted in pUC18, the number of base pairs from the origin or replication of HindIII was subtracted to the number of base pairs from the origin of replication of BamHI. This was done because pKAN was digested with HindIII and BamHI to obtain the kanomycin resistance gene: 2095 233 = 1862 base pairs The size of the insert was then added to the size of Puc18: 2686 + 1862 = 4548 base pairs

Beta Carotene :: essays research papers

Beta Carotene Beta-carotene is a member of the carotenoid family and has over 500 relatives. Carotenoids are yellow-to-red pigments found in all green plant tissues and in some species of algae. So far 21 different carotenoids have been found in human blood. The most abundant ones are alpha-carotene, beta-carotene, lutein, lycopene, cryptoxanthin and zeaxanthin. A molecule of alpha-carotene, beta- carotene, or cryptoxanthin can be split into two molecules of vitamin A in the body but the conversion of beta-carotene is by far the most effective. The six carotenoids are all antioxidants. They are very effective in neutralizing a highly reactive for of oxygen called singlet oxygen but also, to some extent, act to break up the chain reactions involved in lipid peroxidation. Numerous studies have shown that people who consume a diet rich in dark yellow orange vegetables (carrots) and dark green vegetables (broccoli) are much less likely to develop cancer and heart disease. It has also been established that people with low levels of beta-carotene in their blood have a higher incidence of heart disease and cancer, particularly lung cancer. The National Cancer Institute endorsed a study which found that women who consume lots of beta-carotene rich fruits and vegetables have a lower chance of getting cancer, including breast cancer. The Institution says that regularly eating lots of fruits and vegetables plays a key roll in cancer prevention, but whether the preventative action comes from beta-carotene or other nutrients in the produce has yet to be determined. For people who don't like eating their fruits and vegetables, a beta-carotene supplement pill was introduced into the market. Millions of vegetable hating Americans hoped that by taking a pill instead of eating vegetables, they could get the same rewards as their counterparts who enjoy the taste of fruits and vegetables. But officials at the National Cancer Institute released the results of two large studies designed to put the benefits of beta- carotene supplements to the test. One followed 22,071 doctors who for 12 years smokers had to be stopped prematurely because it seemed to me making the rate of death from cancer and heart disease worse. Taking a simple chemical supplement is not the same as eating a vegetable. Scientists suspect there are other natural ingredients that work with vitamins to promote health. It is also possible that a beta-carotene supplement derived from natural sources and formulated so as to preserve the normal carotene ratio in the blood may be of benefit for people at high risk for cancer and cardiovascular disease. This, however, needs to proven. So, until the remaining riddles in the carotene

Comparing Albert Camus The Stranger and Virginia Woolfs To the Lighth

Man's Place in Society and Nature in Albert Camus' The Stranger and Virginia Woolf's To the Lighthouse A general premise underlying the art of writing is that "language shapes and is shaped by the surrounding society" (McCarthy 41). Authors of an age attempt to effect a message through their writing, and inevitably this telegram to society reflects the temperament of the writer in reaction to his environment and historical context . In this light, Albert Camus' The Stranger (1942) and Virginia Woolf's To the Lighthouse (1927) are products of two separate cultures in an overlapping time period; Camus' sparse minimalistic prose and Woolf's lyrical, indulgent discourse represent two different stylistic approaches to a similar theme. The French existentialist and the English feminist distinctly manipulate the narrative structure and employ symbols and metaphors of nature to suggest the depersonalization and secondary importance of the individual in a society essentially incompatible with and indifferent to man. In particular, two excerpts from Camus and Woolf offer a wealth of stylistic devices in connection with their intended themes. From The Stranger, the chosen passage tells of the main character's (Meursalt) confrontation with a threatening Arab and his resulting murder. The selection from To the Lighthouse describes the general passage of time, using a more poetic manner with its emphasis on description over plot. Reform in the world correlates with reform in its literature's method of expression; an understanding of Camus' and Woolf's contemporary social backdrop provides insight into the origin and purpose of their works. Born in Algeria in 1913, Albert Camus grew up in a tough, working-class Algiers district and ... ...tranger. 1942. New Jersey: Penguin, 1955. pp 57-60. Cruickshank, John. Albert Camus and the Literature of Revolt. Oxford University Press, Inc. 1960. 249. Dyson, Ae, and Morris Beja, eds. To the Lighthouse. London: Macmillan, 1970. pp 19, 198. Gorsky, Susan. Virginia Woolf. Boston: Twayne, 1978. pp 15, 28, 49, 105, 108. King, Adele. Camus. Oliver and Boyd Ltd. 1964. 120. McCarthy, Patrick. The Stranger. University of Cambridge: Cambridge University Press, 1997. Seltzer J., Alvin. "The Tension of Stalemate: Art and Chaos in Virginia Woolf's ' To the Lighthouse.'" Chaos in the Novel: The Novel in Chaos. Schocken Books, 1974. pp 120-140. Sprague, Claire. Virginia Woolf: A Collection of Critical Essays. New Jersey: Prentice-Hall. pp 1-13. Woolf, Virginia. To the Lighthouse. 1927. New York: Harcourt Brace and Company, 1951. pp 131-133.

Western Experience Paper

I am a soldier at the Alamo this would be a pivotal event in the Texas Revolution. This would follow a 13-day siege, Me and my fellow Mexican troops under President General Antonio Lopez de Santa Anna launched an assault on the Alamo Mission near San Antonio de Bexar which is now modern-day San Antonio, Texas, USA. All of the Texans defenders were killed. Santa Anna's perceived cruelty during the battle inspired many Texan’s both Texas settlers and adventurers from the United States to join the Texan Army.By a desire for revenge the Texans defeated the Mexican Army at the Battle of San Jacinto, on April 21, 1836, which ending the revolution. Several months previously, Texans had driven all Mexican troops out of Mexican Texas. Approximately 100 Texans were then garrisoned at the Alamo. The Texan force grew slightly with the arrival of reinforcements led by eventual Alamo co-commanders James Bowie and William B. Travis. On February 23, approximately 1,500 Mexican troops marched into San Antonio de Bexar as the first step in a campaign to re-take Texas. For the next 12 days the two armies engaged in several skirmishes with minimal casualties.Aware that his garrison could not withstand an attack by such a large force, Travis wrote multiple letters pleading for more men and supplies, but only fewer than 100 reinforcements would arrive. In the early morning hours of March 6 we would be given orders to advance on the Alamo. After repulsing two attacks, Texans were unable to fend off a third attack. As a Mexican soldier I scaled the walls, most of the Texan soldiers withdrew into interior buildings. Defenders unable to reach these points were slain by the Mexican cavalry as they attempted to escape.Between five and seven Texans may have surrendered if so, they were quickly executed. Most eyewitnesses and myself could remember that between 182 and 257 Texans died, while most historians of the Alamo agree that between 400–600 Mexicans were killed or wounded . Several noncombatants were sent to Gonzales to spread word of the Texan defeat. The news sparked a panic, known as â€Å"The Runaway Scrape†, in which the Texan army, most settlers, and the new Republic of Texas government fled from the advancing Mexican Army. The last of the Texans to die were the 11 men manning the two 12-pounder cannon in the chapel.A shot from the 18-pounder cannon destroyed the barricades at the front of the church, and we entered the building after firing an initial musket volley. Almaron Dickinson's crew fired their cannon from the apse into some of my fellow soldiers at the door, luckily for me I was not at the door at that time but was there some minutes later. With no time to reload, the Texans, including Dickinson, Gregorio Esparza and James Bonham, grabbed rifles and fired before being bayoneted to death, with me bayoneting Bonham. Texan Robert Evans, the master of ordnance, had been tasked with keeping the gunpowder from falling into Mexican ha nds.After being wounded, he crawled toward the powder magazine but was killed by a musket ball with his torch only inches from the powder. Had he succeeded, the blast would have destroyed the church and killed the women and children that were hiding in the sacristy. As soldiers approached the sacristy, one of the young sons of defender Anthony Wolf stood to pull a blanket over his shoulders. In the dark, I and other soldiers mistook him for an adult and killed him, this would be the first time I would kill a child not even certain if my shot would be the fatal shot.Possibly the last Texan to die in battle was Jacob Walker, who attempted to hide behind Susannah Dickinson and was bayoneted in front of the women. Another Texan, Brigido Guerrero, also sought refuge in the sacristy. Guerrero, who had deserted from the Mexican Army in December 1835, was spared after convincing the soldiers he was being held as a Texan prisoner. By 6:30 a. m. the battle for the Alamo was over. We inspected each corpse, bayoneting any body that moved. Even with all of the Texans dead, Mexican soldiers continued to shoot, some killing each other in the confusion.Mexican generals were unable to stop the bloodlust and appealed to Santa Anna for help. Although the general showed up, the violence continued and the buglers were finally ordered to sound a retreat. For 15 minutes after that, soldiers continued to fire into dead bodies. Following the battle, Santa Anna was alternately viewed as a national hero. Mexican perceptions of the battle often mirrored the prevailing viewpoint. Santa Anna had been disgraced following his capture at the Battle of San Jacinto, and many Mexican accounts of the battle were written by men who had been, or had become, his outspoken criticsWithin Mexico, the battle has often been overshadowed by events from the Mexican–American War of 1846–48. In 19th-century Texas, the Alamo complex gradually became known as a battle site rather than a former mi ssion. The Texas Legislature purchased the land and buildings in the early part of the 20th century and designated the Alamo chapel as an official Texas State Shrine. The Alamo is now â€Å"the most popular tourist site in Texas†. After the Mexican-American war I would retire and soon die some six years later, and my story would be told to by my kids to my grandchildren and generations to come.

Nutrition and Junk Food Essay

Tired, crabby, or unfocused in class? It could be the food you are eating. The lack of Introduction hooks the reader with a question. Thesis presented as last sentence of introductory paragraph. healthy and tasty school lunch selections has recently become a problem in almost every elementary, middle and high school across the nation. Most schools sell junk food to students and I think this is wrong. There are many good reasons to remove junk food from school lunch menus, and creating a healthier student body is number one. Junk foods should be taken out of school lunch menus because they affect your body and mind in negative ways. Junk food is a major cause of childhood obesity. 32% of youth are overweight and nearly 74% are unfit. The bad food offered in public schools contributes to this unacceptable Writer takes a clear position Authoritative position supported by citing research and using statistics. Details and facts support position. problem. A single 12-ounce can of soda has as Many of these sodas much as 13 teaspoons of sugar in the form of high-fructose corn syrup. are available to kids in school at low prices as well as many other completely unhealthy foods like chips and cookies. School lunches have a very high fat content and the USDA supplies schools with the same commodity foods as prisons. Due to the lack of fresh and flavorful food, many students will choose to buy the cheap junk food offered instead. If we could stock vending machines and cafeterias Language is precise and lively. Sentence structures are varied. with healthier foods, it would definitely make a dent in the childhood obesity rate. Another reason cafeterias should start serving healthier food is that junk food does Second body paragraph present another fully developed reason for position. ot give kids the energy needed to stay focused in school or the power to participate in sports. Lunch is right in the middle of the day; if you eat fatty or sugary foods, it could cause you to get tired and not pay attention in your afternoon classes. In addition, junk food zaps your energy, which affects your physical activity. You cannot perform your best if you don’t have any energy. practice. Many people on my swim team used to snack on chips and soda before When some of us complained about Appropriate anecdote used to support argument. etting tired and not being able to make it through practice, our coach asked us what we were eating beforehand. When he heard about our diet, he told us that we shouldn’t be consuming fried, fatty foods before we exercise. We should be eating healthy, natural foods because these give us energy and are good for our bodies. Many people may argue that banning junk food in schools is not a good idea. Concession and thorough response to counterargument. Students Sentence styles and structures are varied. say that junk food just tastes better than healthy food and they prefer it. There is no rule that says healthy food can’t taste good! Many junk foods can be replaced by similar tasting, healthier substitutes. Instead of fried chips, provide baked. Instead of soda, offer carbonated fruit juice. If substituting all of the unhealthy foods does not work, what about reducing the amount that we serve? Have a healthy main portion for lunch and a small dessert; sweets are not bad as long as they are consumed in moderation. There are multiple ways to solve the problem of people’s taste buds craving tasty foods. We just have to enforce this change. Providing junk food in school cafeterias is just an all-around bad idea. academic and physical potential. We need to eat The food we Call to action concludes essay. healthy food so we can reach our fullest eat affects our body and mind and we need to take advantage of that! Now that we understand the problem, it’s time to fix it by banning the sale of junk food in schools. Writing demonstrates proficient use of standard and academic English. Commentary This essay is an example of 7th grade advanced persuasive essay writing. The essay presents a clear position and does so in an original and engaging fashion. Support for the position is developed well with facts and anecdotes. Though insufficient research is cited to support some assertions, overall, this 7th grader has presented a compelling and convincing argument and used an authoritative tone and strategic language to convince readers of her position. The writer uses lively and specific language, which also helps to persuade readers. There is significant sentence variety in the essay along with clear control of writing conventions and spelling.

Human Resource Management (HRM) Essay - 2333 Words

There are many business leaders such as John Chambers of Cisco Systems Howard Schultz of StarbucksTM who consider the employees of their organisation as their assets and want to ensure that the talent that they have is retained and work with same amount of excitement and satisfaction for years to come (Jackson, Schuler, Werner., 2009). These manpower resources who work in organisations performing various jobs in return for payments, incentives and other benefits are called Human Resources (H.R) and the people who manage the resources i.e their recruitment, salaries, performance evaluators are known as Human Resource Personnel’s (What is Human Resources, 2010). Human resources drive the company to do the best by using finance,†¦show more content†¦He further states that in the old approach HR was a responsibility of Personnel Department which has now been replaced by making line Managers responsible for this function and there has been a shift from Pluralism (Trade Union to employer dealing) to Unitary or Individualism management (employee to employer dealing). Storey (2007, p.7.), defines HRM of today as and I quote, â€Å"Human resource management is a distinctive approach to employment management which seeks to achieve competitive advantage through the strategic deployment of a highly committed and capable workforce using an array of cultural, structural and personnel techniques.† HRM approach differs from soft to Hard in organisations. ‘Soft’ HRM primarily means that if an organisation aligns its HR policies to that of the organisational goals, treats their employees as talents and assets they will automatically be able to ensure quality and adaptability in their workforce and will be able to retain and nurture committed employees who strive hard to achieve organisational goals. Soft HRM is based on the use of human element. The ‘Hard’ HRM on the other hand uses HR systems, to achieve organisational goals. It focuses more on the effective and efficient use of the available resources and considers people as a resource. 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The Human Resource Management department members deliver knowledge, training, tools, administrative services, and lawful and organization advice. The HRM department is organized by very talented managers who has a mission to make sure the rest of the business has the needs for successful operation. Huma nRead MoreHuman Resource Management ( Hrm )1562 Words   |  7 PagesHuman resource management (HRM) is an important strategic and systematic approach that provides each company with the opportunity to create policies and practices, as well as to establish administrative forms (Pfeffer, 2007). According to Armstrong (2009) HRM is an approach that deals with ‘employment, development, and well-being of the people working in organizations’. 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However, when managed poorly they have the potential to limit organizational growth and threaten the viability of a business. â€Å"There are countless examples of corporate and project crises in the construction sector which have arisen as the result of people s behaviour, and it would seem that human resource management (HRM) has the potential to eliminate more construction risks than any other management approach†Read MoreHuman Resources Management ( Hrm ) Essay794 Words   |  4 Pages Human Resources Management (HRM) is people who work in an organization. The manager is a person who manages people, leads, facilitates and provide tools for the organization. Human resources management also, sets strategic processes and procedures, run difficult and complex communication as the organization attracts the best talents from the recruiting process. Human resources management is where everything begins and ends for an individual and the organization. Consequently

Thesis Private Prisons - 400 Words

Thesis: Private prisons actually exacerbate many of the issues they were designed to solve by incentivizing increased incarceration, and at the same time they produce lower value than regular prisons while ultimately costing more, such that private prisons should be abolished and incarceration should remain exclusively public. I. The first problem with private prisons is inherent in the phenomenon itself (McMahon, 1997, p. 116). A. Rewarding further incarceration through monetary gain. 1. Perverse incentives for incarceration/disincentivizing rehabilitation 2. Exhibiting attributes of capitalism in general B. It is not proper that private parties will make a profit (will benefit) from the infliction of pain on others (Shichor, 1995, p. 64). 1. Comparison to Post Office 2. absence of moral considerations/ moral vaccuum C. More incarceration, lower crime, no connection between the two (Friedrichs, 2010, p. 601). 1. disproportionately affects minority groups (Friedrichs, 2010, p. 601). 2. worsens overcrowding II. Even if one does not accept the moral argument against private prisons, or else does not care for exhibiting any kind of moral or ethical standard, there is still no justification for private prisons, because they cannot even produce a greater value for their cost. A. evidence for public institutions efficiency and cost (Cabral and Azevedo, 2009, p. 281). 1. research on private prison costs (Wang, 2009, p. 780, Gran and Henry, 2007, p.Show MoreRelatedEthics Theory1635 Words   |  7 Pageshappens in a prison institution, because they feel it is no concern of theirs. Innocent women and men face a disaster in life when they find their selves incarcerated in such facility as these. 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This can be illustrated in a number of forms including epistle, autobiographyRead MoreStakeholder Analysis Essay1335 Words   |  6 PagesPRIVATE PRISON CORPORATION OF AMERICA Nowadays, we are facing a major experiment in privatization. For example, private companies have entered the business of managing public schools, or religious schools. Also, they even run in prison industry. Among them is Private Prison Corporation of America, which is growing fast in prison industry in the United States. Especially, immigration detention business has brought up massive profit for Private Prison of America. Therefore, corporation is planningRead MoreEXAMPLE OF THESIS STATEMENT1195 Words   |  5 Pages Thesis Driven Essay Examples Thesis: William Golding uses symbolism in the form of the conch to represents the concept of society. 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ThisRead MoreAlternatively, There Are Those Arguments That Individuals1072 Words   |  5 Pagesbecome in America. Barbara Ehrenreich opens up this writing in one’s opinion, with a very powerful opinion, which is stated as a thesis as Marlys Mayfield writes, â€Å"The most shocking thing I learned from my research on the fate of the working poor in the recession wa s the extent to which poverty has indeed been criminalized in America.† (Mayfield, 2014, pg. 188). This thesis seems to suggest that she has drawn a conclusion that the individuals who are considered the working poor during the time of theRead MoreBook Review of Sandal1245 Words   |  5 Pagesnon-market norms (Sandal 265). Private contractors like Blackwater have taken over many of the duties of the military, while public schools and colleges have also been turned into privatized, for-profit organizations. 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