Constructing a test system for the identification of plague microbe based on genetic probes

• Published in: Genetika Vol. 27; no. 12; p. 2063
• Main Authors: Popov, Iu A, Bulgakova, E G, Kulichenko, A N, Shishkina, O G, Lezhnev, A I, Fedotov, E A, Kirillina, O A
• Format: Journal Article
• Language: Russian
• Published: Russia (Federation) 01-12-1991
• Subjects: Bacteriological Techniques; DNA Probes; DNA, Bacterial - analysis; Plasmids - genetics; Yersinia pestis - isolation & purification
• ISSN: 0016-6758

DNA probes for detection of the plague agent Yersinia pestis were made on a basis of its three typical extrachromosomal replicons. The recombinant plasmid pBS2 including pBR327 vector and SalGI-BspRI fragment of the plasmid pFra was constructed. The above fragment is connected with synthesis of Y. pestis capsular antigen and it is a 400 bp species-specific DNA probe called F1 which is suitable for identification of Y. pestis species that bears the 60 mdal plasmid. The DNA probes called P1 was made on a basis of the plasmid pPst; it is the 460 BglII-BamHI fragment of the fibrinolysin-coagulase gene suitable for species-specific detection of Y. pestis species that bears the 60 mdal plasmid. The P1 fragment was cloned into the pAT153 vector and the constructed recombinant plasmid was called pEK7. The recombinant plasmid pCL1, including the pBR325 vector and the 6th BamHI fragment of Y. pestis EV plasmid pCad was constructed. The above fragment includes the replication origin of the pCad and it is hybridized to the pCad-bearing strains of Y. pestis and Y. tuberculosis only. Thus, it may be a basis for a bi-species-specific DNA probe making. These three recombinant plasmids are considered as a test-system for detection of both typical and atypical strains of Y. pestis.