Gene replacement and retrieval with recombinant M13mp bacteriophages.
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TL;DR: An allele exchange system for shuttling sequences of DNA to and from their original chromosomal loci by exploiting unique properties of M13 lysogens to devise positive selections to detect integration and excision and manipulate phenotypically silent alleles.
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Abstract: We have developed an allele exchange system for shuttling sequences of DNA to and from their original chromosomal loci. Cloned segments of the histidine operon of Salmonella typhimurium and the lactose operon of Escherichia coli served as target sequences and were used to develop the system. Replacement and retrieval of target sequences used the phage M13mp vectors and proceeded through an M13 lysogen intermediate. The intermediates and products of allele exchange were characterized by genetic and hybridization analyses. Several unique properties of M13 lysogens were exploited to devise positive selections to detect integration and excision. These positive selections were used to manipulate phenotypically silent alleles.
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Citations
Increased thermosensitivity associated with topoisomerase I deletion and promoter mutations in Escherichia coli
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23
UV mutagenesis in Salmonella typhimurium is umuDC dependent despite the presence of samAB.
TL;DR: It is found that UV mutability was eliminated when any of three different umuDC alleles were on the chromosome even when samAB was present, which means that samAB normally does not complementUmuDC function in S. typhimurium.
21
Chemotaxis in Escherichia coli proceeds efficiently from different initial tumble frequencies.
TL;DR: The relationships between the level of tumbling, tumble frequency, and chemotactic ability were tested and it appears that a largely smooth-swimming mutant may undergo chemotaxis by increasing tumbling frequency in negative gradients.
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Mutations that affect transcription and cyclic AMP-CRP regulation of the adenylate cyclase gene (cya) of Salmonella typhimurium.
J P Fandl,L K Thorner,S W Artz +2 more
TL;DR: The results establish that the cya gene is transcribed from multiple promoters one of which, P2, is negatively regulated by the cAMP-CRP complex.
17
Amino acid substitution analysis of E. coli thymidylate synthase: the study of a highly conserved region at the N-terminus.
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