9700_m23_qp_42
A paper of Biology, 9700
Questions:
10
Year:
2023
Paper:
4
Variant:
2

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Interferon-alpha (IFN-α) can be produced as a recombinant human protein to treat some types of cancer. The gene IFNA2 codes for IFN-α. One method of producing recombinant IFN-α uses genetically engineered Escherichia coli bacteria that contain recombinant plasmids. Each recombinant plasmid contains: • the gene IFNA2 • three regulatory sequences of the lac operon (promoter, operator and lac• a gene for antibiotic resistance, AMPR. Each of the sequences for the lacI gene and AMPR gene contains its own promoter. As a result, these genes are always expressed in E. coli bacteria that contain this recombinant plasmid. is a diagram of the recombinant plasmid. The promoter regions of the lacI gene and AMPR gene are not shown. IFNA2 AMPR lacI o p e r a t o r p r o m o t e r The start of transcription of the gene IFNA2 by E. coli with the recombinant plasmid shown in needs to be controlled to obtain an optimum yield of IFN-α. Scientists investigated the effect of two inducers of transcription on the production of recombinant IFN-α: • lactose, which is converted to allolactose in E. coli • IPTG, which is a synthetic molecule with a very similar structure to allolactose. IPTG cannot be broken down by E. coli. The scientists grew three cultures of E. coli containing the recombinant plasmid in the same growth medium. The growth medium contained glucose, amino acids, essential vitamins and minerals. The growth medium did not contain lactose. After four hours, either lactose or IPTG at the same concentration was added to two of the cultures of E. coli. As a control, the third culture of E. coli was grown without adding lactose or IPTG. The concentration of recombinant IFN-α in the cultures was measured at different times over a period of 28 hours. The results are shown in . time / hours concentration of IFN-α / μg dm–3 key culture to which IPTG added culture to which lactose added control culture The regulatory sequences of the lac operon contained in the recombinant plasmid are involved in the control of transcription of the gene IFNA2. Explain the role of the gene lacI in the control of transcription of the IFNA2 gene between 0 hours and 4 hours. With reference to , describe the changes in the concentration of recombinant IFN-α in the culture containing IPTG from when IPTG was added at 4 hours to the end of the experiment at 28 hours. Suggest one reason for the difference between the concentration of recombinant IFN-α in the culture at 8 hours in the presence of lactose and the concentration of recombinant IFN-α in the culture at 8 hours in the presence of IPTG. Suggest one reason for the change in the concentration of recombinant IFN-α in the culture containing IPTG from 12 hours to 16 hours. The gene AMPR in the plasmid shown in codes for a protein that provides resistance to the antibiotic ampicillin. Suggest how AMPR allows genetically engineered E. coli containing the recombinant plasmid to be identified. Bacteria can evolve antibiotic resistance through natural processes. Outline how bacteria can evolve to become resistant to antibiotics.
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Salmon can be genetically modified (GM) to produce increased quantities of growth hormone, which is a protein. GM salmon modified in this way have a faster growth rate and reach their maximum body mass at a younger age than non-GM salmon. Within any population of salmon there is variation in body mass. This is an example of continuous variation. Explain what is meant by continuous variation and how it can be caused. Scientists investigated whether injection of very young non-GM salmon with recombinant growth hormone could cause an increase in the growth rate of the salmon. The scientists used two groups of non-GM salmon: • a control group of salmon that were not injected with recombinant growth hormone • an experimental group of salmon that were injected with 1.0 µg of recombinant growth hormone at the start of the experiment and once a week for the next six weeks. The mean body mass of the salmon in the two groups at the start of the experiment was the same (5.3 g). After six weeks, the body mass of every salmon was measured again. The results are summarised in Table 3.1. Table 3.1 no injection with recombinant growth hormone injected with recombinant growth hormone number of non-GM salmon 28 body mass / g range 6.5–8.6 7.2–12.7 mean (xr ) 7.7 9.4 standard deviation 0.4 1.1 A student decided that a t-test should be performed on the results shown in Table 3.1. Calculate the value of t for the results shown in Table 3.1 using the formula for the t-test: t = n s n s x x + - r r f p Give your answer to two decimal places. Show your working. t = The critical value at p = 0.05 for these data is 2.01. The student used the results in Table 3.1 and the t-test to conclude that the injections of recombinant growth hormone cause an increase in the growth rate of the non-GM salmon. Comment on the extent to which the conclusion made by the student can be supported. Suggest one advantage, other than cost, of farming GM salmon that produce increased quantities of growth hormone instead of farming non-GM salmon that are injected with recombinant growth hormone each week.
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Array comparative genome hybridisation (aCGH) is a technique involving the use of a microarray to analyse a genome or sections of a genome. Outline the steps required to prepare the genome of an individual so that the genome is ready for analysis using a microarray chip. DiGeorge syndrome is a dominant inherited disease in humans. DiGeorge syndrome is caused by deletion of a large number of nucleotides from chromosome 22. The number of nucleotides deleted varies between individuals in a range from 800 000 to 3 100 000. The largest deletions can cause the removal of up to 46 protein-coding genes from the chromosome. shows the results of aCGH using a microarray specific for the section of chromosome 22 within which the DiGeorge syndrome deletion occurs. The microarray analysed DNA from two individuals: • one with DiGeorge syndrome • one who did not have DiGeorge syndrome (control DNA for comparison). In the aCGH results shown in : • Each small circle represents the results from a single probe on the microarray. • The x-axis shows the position of each probe on chromosome 22. The position is shown as distance along the chromosome in millions of nucleotides. • A result close to 100% fluorescence on the y-axis means that the DNA from the individual with DiGeorge syndrome fluoresces at the same intensity as the control DNA for that probe. • A result close to 50% fluorescence on the y-axis means that the DNA from the individual with DiGeorge syndrome fluoresces half as much as the control DNA for that probe. 16.0 17.0 18.0 position of probe on chromosome 22 / millions of nucleotides 19.0 20.0 21.0 fluorescence of DNA from an individual with DiGeorge syndrome as a percentage of the fluorescence of control DNA With reference to , estimate the number of nucleotides deleted from the affected chromosome 22 in the individual with DiGeorge syndrome. Give your answer to the nearest 100 000 nucleotides. Explain how the microarray technique works to give the results shown in . Suggest why the phenotypes of two individuals with DiGeorge syndrome can be different.
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