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Identification of a Salmonella typhimurium genomic region involved in invasion of HeLa and Henle‐407 epithelial cells
Author(s) -
Park Jeong U.,
Smith Stuart C.,
Coloe Peter J.
Publication year - 2000
Publication title -
apmis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.909
H-Index - 88
eISSN - 1600-0463
pISSN - 0903-4641
DOI - 10.1034/j.1600-0463.2000.d01-4.x
Subject(s) - cosmid , biology , microbiology and biotechnology , genomic dna , plasmid , hela , genomic library , cloning (programming) , genomic island , salmonella , gene , genetics , cell culture , bacteria , peptide sequence , computer science , programming language
To identify the invasion determinant, a cosmid library was constructed by cloning a genomic library of Salmonella typhimurium 82/6915 into a cosmid vector, pLA2917. A genomic region involved in invasion of cultured HeLa and Henle‐407 cells was subcloned into plasmid pGEM‐7Z. E. coli strain DH1 carrying pSV6235 consisting of a S. typhimurium 4.6 kb genomic region in pGEM‐7Z showed invasion of cultured HeLa and Henle‐407 cells. Nested sequential deletions were introduced into the 4.6 kb genomic region of pSV6235. The E. coli recombinants which contained less than 1.5 kb deletions from the 5′ end ( Sma I site) of the genomic region invaded the cells as effectively as DH1 (pSV6235). The invasion of the recombinants carrying over 2.0 kb deletions from the end of pSV6235 was significantly inactivated compared to DH1 (pSV6235). Restriction enzyme analysis showed that the 3.1 kb fragment from the 3′ end of the 4.6 kb genomic region was distinguished from the Salmonella pathogenicity I genes of S. typhimurium such as the inv, spa , and hil regions showing invasion of the cultured eukaryotic cells.