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Cloning and detailed mapping of the fra-ymt region of the Yersinia pestis pFra plasmid

Cherepanov, P.A.; Mikhaĭlova, T.G.; Karimova, G.A.; Zakharova, N.M.; Ershov, I.V.; Volkovoĭ, K.I.

Molekuliarnaia Genetika Mikrobiologiia i Virusologiia 12: 19-26

1991

ISSN/ISBN: 0208-0613
PMID: 1787840
Document Number: 379008
The genetical libraries of the pFra plasmid of Yersinia pestis genes were obtained by insertion into the PstI, SalGI, EcoRI, XhoI restriction sites of the cosmid vector pHC79. The immunochemical analysis of the recombinant clones has revealed the clones synthesizing the antigen Fl (fraction I) and mouse toxin (Ymt--Yersinia pestis murine toxin). The restriction analysis of the plasmids from antigen synthesizing clones has permitted to construct the detailed physical map of the fra-ymt region of the pFra plasmid the size of 22 kb. The recombinant F1 positive clones of Escherichia coli are able to form at 37 degrees C the capsule-like structure peculiar for Yersinia pestis. The antigen F1 and the mouse toxin were isolated, purified and characterized. The antigen F1 is an 1-2 Md polymer containing a 16 kDa protein subunit. The mouse toxin a 240 kDa protein consisting of 61 kDa subunits. The nucleotide sequence of ymt gene has been defined.

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