Complementation of Mutations in and by DNA Cloned in a Broad-host-range Cosmid Vector Free

Abstract

SUMMARY: A gene library of DNA was constructed in using the broad-host-range cosmid vector pLAFR1. The average insert size was 24-9 kb. From 500 members of the gene library, clones were identified which complemented and Thr mutations in but none which complemented or Leu mutations. Four clones were identified which complemented in Anthranilate synthase activity was detected in a strain only when it harboured a plasmid from one of these clones; activity was repressed when tryptophan was included in the growth medium. Two clones were identified which complemented of A recombinant plasmid from one of these clones also restored some of the nitrogen acquisition functions of and glnL in Expression of several virulence-associated products (haemolysin, heat-labile toxin, adenylate cyclase, filamentous haemagglutinin, and the cell-envelope polypeptide of 30000) was not detected in 500 independent clones. However, by transferring the recombinant plasmids to a mutant of deficient in haemolysin and adenylate cyclase, a plasmid was identified which restored both these activities.

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1986-11-01
2024-03-29
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