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#Question id: 15620
#Unit 13. Methods in Biology
Wild type E. coli metabolizes the sugar lactose by expressing the enzyme ß-galactosidase. You have isolated a mutant that you call lac1–, which cannot synthesize ß-galactosidase and cannot grow on lactose (Lac–). During an condition Lac– strain, called lac3–, is linked to the Tn5 insertion. From a strain carrying the Tn5 insertion and lac3– mutation you isolate an F’ that caries a region of the chromosome that includes both Tn5 and the linked Lac region. Introduce this F’ into an F– strain carrying lac1– by selecting for Kanr. These merodiploids express ß-galactosidase normally. What does this result tell you about the relationship between the lac3– and lac1- mutations?
TLS Online TPP Program
#Question id: 15621
#Unit 13. Methods in Biology
Wild type E. coli metabolizes the sugar lactose by expressing the enzyme ß-galactosidase. You have isolated a mutant that you call lac1–, which cannot synthesize ß-galactosidase and cannot grow on lactose (Lac–). During an condition Lac– strain, called lac3–, is linked to the Tn5 insertion. From a strain carrying the Tn5 insertion and lac3– mutation you isolate an F’ that caries a region of the chromosome that includes both Tn5 and the linked Lac region. Introduce this F’ into an F– strain carrying lac1– by selecting for Kanr. These merodiploids express ß-galactosidase normally. If the merodiploid were Lac–,
a) the two mutations lie in the different gene.
b) Both mutation are complement each other
c) the two mutations lie in the same gene.
d) Both of the mutations is dominant to wild type.
what could you conclude about the relationship between the lac3– and lac1– mutations from these statements?
TLS Online TPP Program
#Question id: 15622
#Unit 13. Methods in Biology
9 In your study of a new bacterial species you have identified a transducing phage that you call Px. In addition you have worked out methods to make random transposon insertions into the bacterial genome. You have generated two different transposon insertion collections one with 105 random Tn5 (Kanr) insertions and the other with 105 random Tn10 (Tetr) insertions. You grow Px phage on the mixed collection of Tn5 insertions and use the resulting phage lysate to infect the mixed collection of Tn10 insertions. You select 10,000 Kanr transductants and find that 80 of them are Tets. Use this information to estimate the total size of the bacterial genome assuming that both Tn5 and Tn10 insert randomly and that the average size of a fragment recombined into the recipient genome during Px transduction is 55 kbp. (Tn5 is about 5 kbp and Tn10 is 10 kbp.)
TLS Online TPP Program
#Question id: 15637
#Unit 13. Methods in Biology
You have isolated two mutations in the Lac operon that cause constitutive expression of Lac genes. You designate these mutants Lac1– and Lac2–. Making use of an F' that carries the Lac operon with the LacY gene mutated, you construct strains that you test for both ß-galactosidase activity and Lac permease activity with results shown below.
Classify each mutation as dominant or recessive and as cis- or trans-acting, giving the experimental result that allows you to arrive at each conclusion. Finally, deduce what type of Lac mutation best fits the properties of Lac 1– and of Lac 2–.
TLS Online TPP Program
#Question id: 15638
#Unit 13. Methods in Biology
You have isolated two mutations that show decreased expression of the Lac operon. However, unlike like the promoter mutations, these mutations don’t respond to the inducer IPTG. These mutations, designated Lac3– and Lac4–, are evaluated for the quantity of ß-galactosidase and permease activity expressed with or without IPTG:
Mapping experiments reveal that Lac3– and Lac4– are different short deletions located in the region before the start of the LacZ gene. Given the data shown above suggest which genetic element(s) in addition to part of the promoter has been deleted in each mutant.
TLS Online TPP Program
#Question id: 15652
#Unit 13. Methods in Biology
You are studying a new strain of E. coli that can utilize the disaccharide melibiose very efficiently. You find that utilization depends on the enzyme melibiase, which is encoded by the gene Mel1. Mel1 is not expressed unless melibiose is present in the growth medium. You have isolated a mutation that causes constitutive melibiase activity, which you designate MelA–. P1 phage mapping experiments using a Tn5 insertion linked to Mel1 show that MelA– is not linked to Mel1. Moreover you find that when an amber suppressor is introduced into a MelA– mutant, normal melibiase regulation is restored. Classify the MelA– mutation in terms of its basic genetic properties,