Involvement of IS26 element in the evolution and dissemination of blaSHV-2a and blaSHV-12.
- Author:
Jungmin KIM
1
;
Haeng Seop SHIN
;
Dong Taek CHO
Author Information
1. Department of Microbiology, College of Medicine, Dankook University, Cheonan, South Korea. dtcho@knu.ac.kr
- Publication Type:Original Article
- MeSH:
beta-Lactamases;
Clone Cells;
DNA;
Klebsiella pneumoniae;
Plasmids;
Polymerase Chain Reaction
- From:Journal of the Korean Society for Microbiology
2000;35(3):263-271
- CountryRepublic of Korea
- Language:Korean
-
Abstract:
A clinical isolate of Klebsiella pneumoniae K7746 produced the extended-spectrum beta-lactamase (ESBL) SHV-12. A 6.6 kb BamHI fragment containing the blaSHV-12 gene of K7746 strain was cloned into pCRScriptCAM vector resulting in the recombinant plasmid p7746-C1. The restriction map of 3.6 kb inserted DNA and sequences immediately surrounding blaSHV-12 of p7746-C1 were homologous to plasmid pMPA2a carrying blaSHV-2a. In addition, both blaSHV-12 and blaSHV-2a were expressed from a common hybrid promoter made of the -35 region derived from the left inverted repeat of IS26 and the -10 region from the blaSHV promoter itself. The results indicate that blaSHV-12 and blaSHV-2a may have evolved from a common ancestor in the sequential order of blaSHV-2a first, followed by blaSHV-12. Furthermore, by the PCR mapping method using primers corresponding to the IS26 and blaSHV, the association between IS26 and blaSHV was studied in 12 clinical isolates carrying blaSHV-2a, 27 clinical isolates carrying blaSHV-12, and 5 reference strains carrying blaSHV-1 to blaSHV-5. All 39 strains carrying blaSHV-2a or blaSHV-12 were positive by the PCR, providing confirmative evidence that IS26 has been involved in the evolution and dissemination of blaSHV-2a and blaSHV-12. But 5 reference strains carrying blaSHV-1 to blaSHV-5 were negative by the PCR. Therefore, we concluded that the molecular evolutionary pathway of blaSHV-2a and blaSHV-12 may be different from that of other blaSHV-ESBL, e.g., blaSHV-2, blaSHV-3, blaSHV-4, and blaSHV-5.