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Cloning and characterisation of the serC and aroA genes of Yersinia enterocolitica, and construction of an aroA mutant.
Gene 1989; 84(1):23-30GENE

Abstract

A gene library of Yersinia enterocolitica 8081 was constructed in the cosmid vector pHC79. Recombinants containing the aroA gene, encoding 5-enolpyruvylshikimate 3-phosphate synthase, were identified by complementation of the aroA mutation in Escherichia coli K-12 strain AB2829. All six recombinant plasmids which complemented aroA also complemented the serC mutation in E. coli K-12 strain KL282. Tn5 mutagenesis suggested serC encoding 3-phosphoserine aminotransferase was the proximal gene in an operon with aroA. The nucleotide sequence of a 3-kb HindII-EcoRV fragment encoding the two genes was determined. The serC and aroA open reading frames contain 362 and 428 codons, respectively, and the deduced amino acid sequences share 78% and 81% homology, respectively, with the corresponding E. coli genes. Sequence inspection revealed no obvious terminators or promoters in the intergenic region. The cloned Y. enterocolitica aroA gene was inactivated in vitro and reintroduced into the parental Y. enterocolitica 8081 strain using the suicide vector pJM703.1. Stable aroA insertion mutants of Y. enterocolitica were isolated.

Authors+Show Affiliations

Department of Molecular Biology, Wellcome Biotech, Beckenham, Kent, U.K.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Comparative Study
Journal Article

Language

eng

PubMed ID

2691337

Citation

O'Gaora, P, et al. "Cloning and Characterisation of the serC and aroA Genes of Yersinia Enterocolitica, and Construction of an aroA Mutant." Gene, vol. 84, no. 1, 1989, pp. 23-30.
O'Gaora P, Maskel D, Coleman D, et al. Cloning and characterisation of the serC and aroA genes of Yersinia enterocolitica, and construction of an aroA mutant. Gene. 1989;84(1):23-30.
O'Gaora, P., Maskel, D., Coleman, D., Cafferkey, M., & Dougan, G. (1989). Cloning and characterisation of the serC and aroA genes of Yersinia enterocolitica, and construction of an aroA mutant. Gene, 84(1), pp. 23-30.
O'Gaora P, et al. Cloning and Characterisation of the serC and aroA Genes of Yersinia Enterocolitica, and Construction of an aroA Mutant. Gene. 1989 Dec 7;84(1):23-30. PubMed PMID: 2691337.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Cloning and characterisation of the serC and aroA genes of Yersinia enterocolitica, and construction of an aroA mutant. AU - O'Gaora,P, AU - Maskel,D, AU - Coleman,D, AU - Cafferkey,M, AU - Dougan,G, PY - 1989/12/7/pubmed PY - 1989/12/7/medline PY - 1989/12/7/entrez SP - 23 EP - 30 JF - Gene JO - Gene VL - 84 IS - 1 N2 - A gene library of Yersinia enterocolitica 8081 was constructed in the cosmid vector pHC79. Recombinants containing the aroA gene, encoding 5-enolpyruvylshikimate 3-phosphate synthase, were identified by complementation of the aroA mutation in Escherichia coli K-12 strain AB2829. All six recombinant plasmids which complemented aroA also complemented the serC mutation in E. coli K-12 strain KL282. Tn5 mutagenesis suggested serC encoding 3-phosphoserine aminotransferase was the proximal gene in an operon with aroA. The nucleotide sequence of a 3-kb HindII-EcoRV fragment encoding the two genes was determined. The serC and aroA open reading frames contain 362 and 428 codons, respectively, and the deduced amino acid sequences share 78% and 81% homology, respectively, with the corresponding E. coli genes. Sequence inspection revealed no obvious terminators or promoters in the intergenic region. The cloned Y. enterocolitica aroA gene was inactivated in vitro and reintroduced into the parental Y. enterocolitica 8081 strain using the suicide vector pJM703.1. Stable aroA insertion mutants of Y. enterocolitica were isolated. SN - 0378-1119 UR - https://www.unboundmedicine.com/medline/citation/2691337/Cloning_and_characterisation_of_the_serC_and_aroA_genes_of_Yersinia_enterocolitica_and_construction_of_an_aroA_mutant_ L2 - https://linkinghub.elsevier.com/retrieve/pii/0378-1119(89)90135-2 DB - PRIME DP - Unbound Medicine ER -