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. 1987 Feb;31(2):286–291. doi: 10.1128/aac.31.2.286

Cloning and expression of genes responsible for altered penicillin-binding proteins 3a and 3b in Haemophilus influenzae.

F Malouin, A B Schryvers, L E Bryan
PMCID: PMC174707  PMID: 3551833

Abstract

A Haemophilus influenzae strain (T-1,3) possessing clinical beta-lactam resistance due to altered penicillin-binding protein 3 was used to construct a recombinant cosmid gene bank in Escherichia coli. Three of the recombinant cosmids were capable of transforming a susceptible H. influenzae strain (Rdnov) simultaneously to moxalactam resistance and altered the binding of penicillin-binding proteins 3a and 3b to [35S]penicillin G. Restriction endonuclease mapping of one of the recombinant cosmids, pLB100, was performed to facilitate subsequent subcloning of the gene(s) responsible for the altered penicillin-binding protein 3 (a and b) binding phenotype. Subcloning of individual fragments derived from pLB100 indicated that two adjacent fragments of DNA were both capable of transforming a susceptible Haemophilus strain to moxalactam resistance and altered penicillin-binding protein 3 binding. Expression of plasmid-coded proteins in minicells indicated that one fragment coded for a major 55,000-molecular-weight polypeptide and that the second contained a C-terminal coding region that expressed a 28,000-molecular-weight polypeptide when fused to the N-terminal region of the tetracycline resistance gene. Initial attempts at labeling the plasmid-coded proteins expressed in minicells with [35S]penicillin G were unsuccessful.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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