Microbiology: An Evolving Science (Fourth Edition)
4th Edition
ISBN: 9780393615098
Author: John W. Foster, Joan L. Slonczewski
Publisher: W. W. Norton & Company
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Chapter 12.1, Problem 1TQ
Summary Introduction
To review:
The modification of mutagenesis with transposons to identify mutants defective in the synthesis of amino acid.
Introduction:
Transposons are the segments of DNA (deoxyribonucleic acid) that can change its site within the genome. They can also alter the genetic information possessed within the genome. This property of transposons can be used to transfer the foreign gene into the host. The process by which the genes are transferred within the chromosome of the host organism using transposons is called transposon mutagenesis. This leads to modification or interruption in the function of the original gene.
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A classic way to isolate thymidylate synthase-negative mutants of
bacteria is to treat a growing culture with thymidine and trimetho-
prim. Most of the cells are killed, and the survivors are greatly
enriched in thymidylate synthase-negative mutants.
(a) What phenotype would allow you to identify these mutants?
(b) What is the biochemical rationale for the selection? (That is, why
are the mutants not killed under these conditions?)
(c) How would the procedure need to be modified to select mam-
malian cell mutants defective in thymidylate synthase?
A classic way to isolate thymidylate synthase-negative mutants of bacteria
is to treat a growing culture with thymidine and trimethoprim. Most of
the cells are killed, and the survivors are greatly enriched in thymidylate
synthase-negative mutants.
(a) What phenotype would allow you to identify these mutants?
(b) What is the biochemical rationale for the selection? (That is, why are the
mutants not killed under these conditions?)
(c) How would the procedure need to be modified to select mammalian cell
mutants defective in thymidylate synthase?
In the procedure shown, why was it necessary to link thecoding sequence for the A or B chains to the sequence forβ-galactosidase? How were the A or B chains separated fromβ-galactosidase after the fusion protein was synthesized in E. coli?
Chapter 12 Solutions
Microbiology: An Evolving Science (Fourth Edition)
Ch. 12.1 - Prob. 1TQCh. 12.1 - Prob. 2TQCh. 12.1 - Prob. 3TQCh. 12.1 - Prob. 4TQCh. 12.2 - Prob. 1TQCh. 12.2 - Prob. 2TQCh. 12.2 - Prob. 3TQCh. 12.3 - Prob. 1TQCh. 12.4 - Prob. 1TQCh. 12.4 - Prob. 2TQ
Ch. 12.5 - Prob. 1TQCh. 12 - Prob. 1RQCh. 12 - Prob. 2RQCh. 12 - Prob. 3RQCh. 12 - Prob. 4RQCh. 12 - Prob. 5RQCh. 12 - Prob. 6RQCh. 12 - Prob. 7RQCh. 12 - Prob. 8RQCh. 12 - Prob. 9RQCh. 12 - Prob. 10RQCh. 12 - Prob. 11RQCh. 12 - Prob. 12RQCh. 12 - Prob. 13RQCh. 12 - Prob. 14RQCh. 12 - Prob. 15RQCh. 12 - Prob. 1TQCh. 12 - Prob. 2TQCh. 12 - Prob. 3TQ
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- how will mutation (with - ) will affect E.coli grown in lactose medium. what is the implication of the following genotypes? i+ p+ o+ z- y+arrow_forwardFrom one Escherichia coli strain, five Hfr strains were isolated. The location and orientation of the origin of transfer of each Hfr strain are shown in Figure 1. want to use these five strains to map the locus responsible for thiamine synthesis, called thi. Each Hfr strain is sensitive to rifampicin (RifS) and Thi+. Conjugation experiments are performed between each of the Hfr strains and an F- RifR Thi 9 leu 10 20 30 nadD pyrC trp 40 his 50 60 70 cysG 80 90 metA 100 Hfr1 Hfr2 Hfr3 Hfr4 Hfr5 Figure 1: Chromosome map of Escherichia coli. Five Hfr strains (Hfr1 to Hfr5) were isolated and the location and orientation of the origin of transfer is shown by the arrows in each Hfr strain. Distances in minutes are indicated. Leu: leucine biosynthesis; nadD: NAD biosynthesis; pyrC: pyrimidine biosynthesis; trp: tryptophan biosynthesis; his: histidine biosynthesis; cysG: cysteine biosynthesis; metA: biosynthesis of methionine. 1) What is the selection medium used in these conjugation…arrow_forwardFrom one Escherichia coli strain, five Hfr strains were isolated. The location and orientation of the origin of transfer of each Hfr strain are shown in Figure 1. want to use these five strains to map the locus responsible for thiamine synthesis, called thi. Each Hfr strain is sensitive to rifampicin (RifS) and Thi+. Conjugation experiments are performed between each of the Hfr strains and an F- RifR Thi leu 10 20 T nadD pyrC trp 30 his Donor strain Hfr1 Hfr2 Hfr3 Hfr4 Hfr5 The results are shown in the following table 60 70 cysG Colonies Thi 1000 0 400 0 25 80 90 metA 1,00 Hfr1 Hfr2 Figure 1: Chromosome map of Escherichia coli. Five Hfr strains (Hfr1 to Hfr5) were isolated and the location and orientation of the origin of transfer is shown by the arrows in each Hfr strain. Distances in minutes are indicated. Hfr 3 Leu: leucine biosynthesis; nadD: NAD biosynthesis; pyrC: pyrimidine biosynthesis; trp: tryptophan biosynthesis; his: histidine biosynthesis; cysG: cysteine biosynthesis; metA…arrow_forward
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