9. What are the primary differences between site-specific recombination and general recombination? Give an example of the biological context in which you might expect to find each of these. (One example for each is sufficient.)
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Q: describe Site-Specific Recombination
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Recombination is the process of reshuffling genetic material to create new combinations of genes leading to genetic diversity.
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- 35. M Heinz Shuster collected the following data on the base composition Move ANALYSIS ribgrass mosaic virus (H. Shuster, in The Nucleic Acids: Chemistry and Biology, vol. 3, E. Chargaff and J. N. Davidson, Eds. New York: Academic Press, 1955). On the basis of this information, is the hereditary information of the ribgrass mosaic virus RNA or DNA? Is it likely to be single stranded or double stranded? Percentage A T U Ribgrass 29.3 25.8 18.0 0.0 27.0 mosaic virus Leibniz Institute for Age Research, Fritz Lipmann-Institute. Ribgrass mosaic virus.618. If 32P is used to label both strands of double-stranded DNA, after two rounds of replication, four double-stranded molecules of DNA are produced. Of these,a. one of the four double-stranded molecules of DNA will contain all the 32Pb. two of the four double-stranded molecules of DNA will contain 32P, each with half of the 32Pc. three of the four double-stranded molecules of DNA will contain 32P, each with 1/3 of the 32Pd. all four double-stranded molecules of DNA will contain 32P, each with ¼ of the 32Pe. none of the double-stranded molecules of DNA will contain 32P
- 2. Dr. Kim at Ewha Research Center performed shotgun Sanger sequencing on an unknown DNA sample, and obtained the following reads (12 reads). Since the length of each read is quite short, Dr. Kim ran the original sample in a gel electrophoresis, and realized that the original DNA is just 50 base pairs long. (Please note that the resolution of gel electrophoresis is not so good. Thus, we cannot figure out the exact length of DNA using gel electrophoresis in the real world.) 1) САСТС ССAGT GTACC T 3) GGAGT CAАТC GC 5) GGCTG TGCTT GG 7) GATGG CTGTG 9) CAGTG TAССT GCA 11) TGCAA GCCGA G 2) AGCCG AGATG GCTG 4) CTGCA AGCCG A 6) CTTGG AGTCA ATCGC 8) САСТС ССAG 10) GCTGT GСТTG G 12) TGCTT GGAGT (a) Find the sequence of the original DNA (reconstruction), and align each read with the reconstructed DNA sequence. (hint: put all reads on a ppt slide, and move them around to find overlaps.) (b) Calculate coverage at each nucleotide position of the reconstructed DNA, i.e., how many reads cover that…1. Drag the red box to indicate the restriction site in the following sequence. 5'-... АТА ТСА ТСС TGT GCT GCC T С АТC TСT сTG CCC ...-3' Attempts left: unlimited Check answer 2. Drag the green box to indicate the single base change or SNP in sequence 2. 1) 5-.. АТА ТСА ТСС TGT GCT GCС ТC АТC ТCT GTG CCС ...-3' 2) 5-.. АТА ТСА ТСС TGT GTT GCс ТтC АТС ТCT GTG CCС ...-3'3. A long DNA will be divided into small fragments, and one of them will be attached to the bead and amplified in Ion Torrent sequencing. The beads with different kinds of fragments will be loaded into different wells. The following graphs show the first forty (40) signals from two different wells. The value of one (1) in the y-axis of the graph corresponds to incorporation of one (1) nucleotide to every DNA strand on a bead. (a) You can observe that in every single step the signals are not the multiples of an integer. Explain why such signals are produced and how we can interpret the signals. (b) Using the first forty (40) signals shown below, please figure out the DNA sequence of the fragment attached to the bead in each well. (c) Based on the sequences of the fragments, can we guess the partial sequence of the original DNA sequence (before fragmentation)? If so, what could it be? (d) In the well #1, when the non-zero signals will show up after the 40 steps? Answer this by the step…
- 1) Using the diagram below, sketch in the pattern of bands you would expect to see after digesting the DNA of the TT, Tt and tt genotypes of the TAS2R38 gene. Use the Base Pair Standards on the left of the diagram as a reference in drawing the positions of the bands. Base Pair TASTER GENO TYPE Standards (Base Pairs) TT Tt tt 500 400 300 200 100 501. What structures are these DNA strand likely to adopt in solution (assuming sufficient salt concentration to permit any hybridisation as appropriate)? Draw your answers depicting the hybridisation. a) GCC TTG AGC TTT TTT GCT CAA GGC (b) ATG ACT CTC GAG AGT CAT TTA TTA3. In the following drawing, the top strand is the template DNA, and the bottom strand shows the lagging strand prior to the action of DNA Polymerase I. Three Okazaki fragments are shown and the RNA primers (asterisks) have not yet been removed. 3'- -5' 5********** RNA primer Left Okazaki fragment ↑ RNA primer ↑ -||- Middle Okazaki fragment -||- *********** RNA primer Right Okazaki fragment -3' A. Which Okazaki fragment was made first, the one on the left or the one on the right? B. Which RNA primer would be the first one to be removed by DNA polymerase I, the primer on the left or the primer on the right? For this primer to be removed by DNA polymerase I and for the gap to be filled in, is it necessary for the Okazaki fragment in the middle to have already been synthesized? Explain why. C. Let's consider how DNA ligase connects the left Okazaki fragment with the middle Okazaki fragment. After DNA polymerase I removes the middle RNA primer and fills in the gap with DNA, where does DNA…
- . A closed circular supercoiled DNA is relaxed by treatment with topoisomerase. No matter how much enzyme is used, or how long the experiment is run, the experimenter always finds a gel elec- trophoresis pattern indicating some DNA with one, two, and three superhelical turns in addition to the relaxed (nicked) circle (see fig- ure). Suggest an explanation for this observation. Nicked circle +39. How can we visualize the effect of supercoiling on the structure of DNA? (describe supercoiling)3. Your PhD thesis advisor has given you the task of preparing a human genomic DNA library. a. How will you prepare DNA fragments from the human genomic DNA for use in construction of this library assuming that you want the insert sizes to be about 20000bp in size? b. What vector will you use for construction of this library? c. How will you ligate the genomic DNA fragments into the vector? d. What host will you use for propagation of your library? e. What probe would you use for screening your library assuming that you wanted to isolate the human insulin gene? f. How many clones would you have to screen to be pretty sure that you will indeed find a clone for insulin?