4. With the use of P22 as a generalized transducing phage grown on a pur+ pro* his+ bacterial donor, a recipient strain of genotype pur pro his was infected and incubated. Afterward, transductants for pur+, pro*, and his* were selected individually in experiments I, II, and III, respectively. a. What medium is used in each of these selection experiments? b. The transductants were examined for the presence of unselected donor markers, with the following results: I II III prohis 86% pur his 44% pur pro 20% pro+ his 0% purt his 0% pur pro 14% pro his+ 10% pur his 54% pur prot 61% pro+ hist 4% purt hist 2% pur prot 5%
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- 1. In terms of Spo11, what does it mean if twice as many genetic sequence reads map to one location than another? Explain. 2. What does it mean if genetic reads map in different locations in difgerent strains? Explain in terms of recombination hotspots.35. In five Hfr strains, each of which was used to build a time-of-entry map, the genes entered the recipient cells as follows: Strain 1: N O S S N A Strain 2: A P T H A N Strain 3: P T H A N O Strain 4: A N O S S N Strain 5: N A P T H A Build the map from this data and enter your result.1. A set of 4 Hfr bacterial strains were used in a time-of-entry experiment to map the locations of nine genes (A, B, C, D, E, F, G, H, I). The experiment was terminated after 22 minutes, which did not provide enough time to map all 9 genes with a single Hfr strain. Using the data on the amount of time it took to transfer each gene to a recipient during conjugation (see table below), draw a circular map of the genes on the bacterial chromosome. Your map should also indicate the Hfr origins of transfer with arrows. You don't need to indicate relative distances between genes, but you should use the given times of entry to figure out relative gene order. G H n/a 13 n/a 9 в с n/a n/a n/a |13 n/a 17 n/a 14 n/a n/a| 12 А D E F I Hfr 1 Hfr 2 1 22 10 n/a n/a n/a 2 n/a n/a |8 n/a n/a n/a 11 Hfr 3 n/a 4 Hfr 4 n/a n/a 2 18
- blaA ori lacl Plac rrgB rrgA lacZYA Recombinant Plasmid "B"a. Bacteriophage P22 was used in generalized transduction experiments to infect the Salmonella typhimurium donor strains described in the table below. The resulting phage lysates were then used to infect the S. typhimurium recipient strains listed in the table. In each cross, a phenotype was selected for one of the three genetic markers studied (str, aceA, thrA), and then replicates were performed to select the corresponding recombinants for the other two markers. The results are given in the following table: Recipient strain Selected phenotype Selected recombinants Donor strain str thrA aceAt thrA str aceA+ strs thrA+ aceA thrA+ str aceA Str Ace+ Str ThrA ThrA+ ThrA ThrA+ Ace Ace+ str: gene involved in streptomycin resistance, aceA: gene involved in the use of acetate as a carbon source, thrA: gene involved in the biosynthesis of threonine. Number 60 40 95 5 10 90 What are the selective media used in these three transduction experiments, on the one hand to obtain the selected…The main reason/reasons that motivated the geneticengineers to use E. coli in the production of monoclonal antibodies compared to the usual techniques is/are: O a. The quality of monoclonal antibody produced by the usual method is poor. O b. The establishment of hybridomas is not easy. O. Theusual technique produces only polyclonal antibodies. O d. The isolation of lymphocytes is difficult. O e. The isolation of spleen cells is difficult.
- A B Figure 1 The postgraduate student, Vanessa, cloned her gene of interest into two different vectors; pUC18 and pZERO®-1, and transformed the competent cells with the recombinant vectors. After that, she plated them on the antibiotic plates and incubated her samples for 18 hours. The next day, she realized that the plates were not labelled. (i) Based on the observation above, can you identify which plate is carrying the recombinant pUC18 and pZErOⓇ-1, respectively? (ii) Why are there small colonies surrounding the big colonies in Figure 1 (A)? Explain your answer in detail. (iii) Do you think all the colonies present on the plates in Figure 1 (B) contain the gene of interest? Describe the selection procedures for recombinant pZErO®-1.The following recombinants are recovered when conjugation occurs between an a+d+g+ donor and an a d'g recipient. at dt g+= 84% ad g+= 6% at dgt = 10% ad g+ = less than 1% Which gene is in the middle? a The genes are located too far apart on the bacterial genome to determine which gene is in the middle. d Insufficient data are provided to answer this question.Shown below are the complementation test results involving 4 independently isolated lethal mutants in a bacteriophage. Complementation was assayed by simultaneouly infecting bacteria with two phage strains, each with a different mutation, neither of which could alone lyse the cells. In the table below, a "+" indicates the strains complemented each other and therefore lysed open the bacteria. A "0" indicates no complementation and therefore no cell lysis occurred. Test pair Results 1___2___3___4 1,2 + 1 0 + + 0 1,3 + 2 0 + + 1,4 0 3 0 + 2,3 + 4 0 2,4 + 3,4 + How many genes are there? a. 3 b.1 c. 2 d. 4
- Shown below are the complementation test results involving 4 independently isolated lethal mutants in a bacteriophage. Complementation was assayed by simultaneouly infecting bacteria with two phage strains, each with a different mutation, neither of which could alone lyse the cells. In the table below, a "+" indicates the strains complemented each other and therefore lysed open the bacteria. A "0" indicates no complementation and therefore no cell lysis occurred. Test pair Results 1___2___3___4 1,2 + 1 0 + + 0 1,3 + 2 0 + + 1,4 0 3 0 + 2,3 + 4 0 2,4 + 3,4 + Which mutants are in the same gene? . a. 2, & 3 b.1, 2, 3 & 4 c.1 & 4 d.1, 2 & 4Comment on the failure of p1vir transduction experiments and give a reason why you think the transductions was unsuccessful for some genes. Suggest any alternative methods that could be used to create the mutations that were unsuccessful in these experiments? There are no colonies on the plate for some of the genes. and in DNA sequence, some colonies don't have primers present. For PCR, some colonies are not replaced by kanamycin or tetracycline.You have conducted a transposon mutagenesis experiment using the same PRL27 system that was used during your lab exercise. After allowing for conjugation. you plate the conjugation mix on Luria agar and Luria + Kanamycin agar. After incubation, you count the following number of colonies on each type of plate: Media Dilution Number of Colonies Too Many to Luria 10-5 Count 10-6 173 10-7 28 Luria + Kanamycin 100 324 10-1 217 10-2 3 Determine the transformation efficiency. Note: You can enter you answer using long form (e.g. 1500000) or scientific notation using e notation (e.g. 1.5e6). Do not enter units.