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Cloning and expression in Escherichia coli of three Rickettsia prowazekii genes, coding outer membrane proteins

Emel'ianov, V.V.; Balaeva, N.M.

Molekuliarnaia Biologiia 26(4): 847-858

1992

ISSN/ISBN: 0026-8984
PMID: 1279381
Document Number: 389221
Rickettsia prowazekii (virulent Breinl strain) random genomic DNA fragments were cloned in the lambda gt11 expression vector by using non-palindromic adaptors. Several immunoreactive clones were selected after screening 20,000 individual recombinant plaques with human convalescent serum. Some recombinants synthesized the complete 60 K protein, and others synthesized beta-galactosidase fusion polypeptides containing epitopes of 134 K protein of the R. prowazekii outer membrane. The amplified genomic library was screened with monospecific antibodies directed against abundant 31 K and 29.5 K outer membrane proteins. Several recombinant clones expressing full or part of 29.5 K polypeptide, and none expressing 31 K polypeptide were revealed. The serum of a patient convalescing from epidemic typhus did not react in western blot with recombinant 29.5 K protein.

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