OHIO-1 BETA-LACTAMASE IS PART OF THE SHV-1 FAMILY

被引:30
作者
SHLAES, DM
CURRIEMCCUMBER, C
HULL, A
BEHLAU, I
KRON, M
机构
[1] VET ADM MED CTR, RES SERV, CLEVELAND, OH 44106 USA
[2] CASE WESTERN RESERVE UNIV, DEPT MED, CLEVELAND, OH 44106 USA
[3] CASE WESTERN RESERVE UNIV, DIV GEOG MED, CLEVELAND, OH 44106 USA
关键词
D O I
10.1128/AAC.34.8.1570
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
The OHIO-1 β-lactamase gene was subcloned in a 1.16-kilobase TaqI fragment in the 2.4-kilobase chimeric plasmid pSK04. After directional subcloning into M13 the DNA sequence of this fragment was determined. The results showed an open reading frame of 858 base pairs (bp) encoding a protein of 286 amino acids. The structural gene showed 85, 87, and 60% DNA sequence identity with SHV-1, LEN-1, and TEM-1, respectively, and 93, 85, and 62% predicted amino acid sequence identity, respectively. The 87 bp upstream of the OHIO-1 structural gene had 96% identity with the upstream flanking sequence of SHV-1, including the -35 and -10 consensus sequences and the putative ribosomal binding site. A 223-bp DNA probe derived from a PstI-HaeII fragment in the C-terminal sequence of OHIO-1 had predicted 96, 88, and 61% sequence identity with SHV-1, LEN-1, and TEM-1, respectively. This probe hybridized to SHV-1 and poorly to LEN-1, but not to TEM-1 or a variety of other plasmid-mediated β-lactamase genes, under stringent conditions. Screening of plasmid DNA derived from 40 ampicillin-resistant clinical isolates by Southern hybridization with the 223-bp probe uncovered no strains encoding OHIO-1. Isoelectric focusing of the same collection did identify two strains producing enzymes resembling SHV-1, however. We have also performed a kinetic comparison of OHOI-1, SHV-1, and TEM-1. OHIO-1 and SHV-1 were indistinguishable from each other but could be distinguished from TEM-1. These data clearly place OHIO-1 within the SHV-1 family of β-lactamases.
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页码:1570 / 1576
页数:7
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