TLS Online TPP Program
More Questions
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,
TLS Online TPP Program
#Question id: 15653
#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 on the basis of genetic properties, make a proposal for the type of regulatory functions affected by the MelA– mutation?
TLS Online TPP Program
#Question id: 15654
#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. Next you isolate a mutation, designated MelB–, which gives uninducible melibiase activity. Mapping experiments show that MelB– is linked to Mel1. Using an F' factor that carries the chromosomal region surrounding Mel1, you perform the following genetic tests:
Describe the basic genetic properties of the MelB– mutation?