BIOLOGY 12E CONNECT ACCESS CARD
BIOLOGY 12E CONNECT ACCESS CARD
12th Edition
ISBN: 9781264938513
Author: Raven
Publisher: MCG
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Chapter 16, Problem 1S

You have isolated a series of mutants affecting regulation of the lac operon. All of these are constitutive, that is, they express the lac operon all the time. You also have both mutant and wild-type alleles for each mutant in all combinations, and on F′ plasmids, which can be introduced into cells to make the cell diploid for the relevant genes. How would you use these tools to determine which mutants affect DNA binding sites on DNA, and which affect proteins that bind to DNA?

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Suppose you have six strains of E. coli. One is wildtype, and each of the other five has a single one of thefollowing mutations: lacZ−, lacY−, lacI−, oc, andlacIS. For each of these six strains, describe thephenotype you would observe using the following assays. [Notes: (1) IPTG is a colorless synthetic molecule that acts as an inducer of lac operon expressionbut cannot serve as a carbon source for bacterialgrowth because it cannot be cleaved byβ-galactosidase; (2) X-gal cannot serve as a carbonsource for growth; (3) E. coli requires active lactosepermease (the product of lacY) to allow lactose,X-gal, or IPTG into the cells.] Colony color in medium containing glycerol as theonly carbon source and X-gal, but no IPTG.d. Colony color in medium containing high levels ofglucose as the only carbon source, X-gal, andIPTG.e. Colony color in medium containing high levels ofglucose as the only carbon source and X-gal, butno IPTG
Strain ROFL4 has a premature stop mutation in the lacZ gene, resulting in a nonfunctional b-galactosidase. Otherwise all other parts of the operon are functional. Circle the least number of components for an F' plasmid that will restore normal regulation and function of the lac operon in the resulting partial diploid. (may need more than one) (a) lacI+   (b) lacO+   (c) lacP+   (d) lacZ+   (e) None, cannot be restored.
The following shows the genotype of a partial diploid bacterial cell - where one chromosomal region containing the lac operon in E,coli is given, and the other fragment is from a plasmid carrying another lac operon from another source. The two are separated by a slash (/). The possible answers indicate with a ʺ+ʺ or a ʺ-ʺ whether β-galactosidase would be expected to be produced at induced levels under two circumstances: 1) first in the absence of lactose and 2) second in the presence of lactose. (Assume that glucose is not present in the medium.)Genotype F: I+ Oc Z-/ Fʹ I- O+ Z+   KEY:I+ = wild-type repressorI- = mutant repressor (unable to bind to the operator)Is = mutant repressor (insensitive to lactose)O+ = wild-type operatorOc = constitutive operator (insensitive to repressor)

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BIOLOGY 12E CONNECT ACCESS CARD

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Bacterial Genomics and Metagenomics; Author: Quadram Institute;https://www.youtube.com/watch?v=_6IdVTAFXoU;License: Standard youtube license