[Cloning and expression in Escherichia coli of three Rickettsia prowazekii genes, coding outer membrane proteins].
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.
D000907:Antibodies, Bacterial / Q000276:immunology; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics*; D010582:Bacteriophage lambda / Q000235:genetics; D015153:Blotting, Western; D003001:Cloning, Molecular; D004269:DNA, Bacterial; D000939:Epitopes / Q000276:immunology; D004926:Escherichia coli / Q000235:genetics*; D015870:Gene Expression; D005798:Genes, Bacterial; D005822:Genetic Vectors; D006801:Humans; D011993:Recombinant Fusion Proteins / Q000235:genetics; D012283:Rickettsia prowazekii / Q000235:genetics* / Q000276:immunology; D014437:Typhus, Endemic Flea-Borne / Q000097:blood / Q000276:immunology; D001616:beta-Galactosidase / Q000235:genetics
D000907:Antibodies, Bacterial; D001425:Bacterial Outer Membrane Proteins; D004269:DNA, Bacterial; D000939:Epitopes; D011993:Recombinant Fusion Proteins; D001616:beta-Galactosidase
Klonirovanie i ékspressiia v Escherichia coli trekh genov Rickettsia prowazekii, kodiruiushchikh belki vneshneĭ membrany.