Determination of Enterococcus faecalis groESL Full-Length Sequence and Application for Species Identification
Determination of Enterococcus faecalis groESL Full-Length Sequence and Application for Species Identification
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DOI：
10.1128/jcm.39.9.3326-3331.2001
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发表时间：
2001-09
影响因子：
9.4
通讯作者：
L. Teng;P. Hsueh;Yi-hui Wang;Hsiao-Mann Lin;K. Luh;S. Ho
中科院分区：
文献类型：
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作者：
L. Teng;P. Hsueh;Yi-hui Wang;Hsiao-Mann Lin;K. Luh;S. Ho
部分Cpn60（或GroEL）基因片段的扩增已被用于许多细菌的鉴定，包括肠球菌。为了获得更多粪肠球菌groESL基因的序列数据，测定了粪肠球菌groESL基因的全长序列，其中groES （285 bp）、spacer （57 bp）和groel （1626 bp）。通过GenBank数据库检索发现，推导出的粪肠杆菌GroES和GroEL蛋白与其他细菌的GroES和GroEL蛋白具有显著的同源性。GroEL的用法和样例：粪孢菌与肺炎链球菌的同源性最高（氨基酸序列同源性为81.8%，核苷酸序列同源性为73.0%），其次是玉米乳球菌，而GroES （GroES）与玉米乳杆菌的同源性为60.2%(64.6%)，与乳酸乳球菌的同源性为58.5%(66.2%)，与枯草芽孢杆菌的同源性为57.0%（65.5%）。根据groES序列，anE。建立了粪肠球菌特异性PCR检测，该PCR检测对所有粪肠球菌菌株均呈阳性。以groES或groel为探针的斑点杂交结果表明，这两种基因均可作为粪肠杆菌鉴定的合适靶点。此外，设计了groESL的宽范围pcr -限制性片段长度多态性，以区分8种常见的肠球菌。根据haeiii和RsaI产生的限制性结构可以很容易地区分参考菌株的肠球菌种类。本研究中开发的基于dna的检测方法为临床重要肠球菌物种的鉴定提供了一种替代方法。
ABSTRACT Amplification of the partial Cpn60 (or GroEL) gene segment has been used for identification of many bacteria, includingEnterococcus species. To obtain more sequence data fromgroESL genes of Enterococcus faecalis, the full-length sequence of the E. faecalis groESL genes containing groES (285 bp), spacer (57 bp), andgroEL (1,626 bp) was determined. A database search of GenBank revealed that the deduced E. faecalis GroES and GroEL proteins show significant homology to the GroES and GroEL proteins of other bacteria. The GroEL (groEL) ofE. faecalis had the highest identity withStreptococcus pneumoniae (81.8% amino acid sequence identity and 73.0% nucleotide sequence identity), followed byLactococcus zeae, while GroES (groES) had 60.2% (64.6%) identity with Lactobacillus zeae and 58.5% (66.2%) identity with Lactococcus lactis, followed by 57.0% (65.5%) identity with Bacillus subtilis. Based on the groES sequence, anE. faecalis-specific PCR assay was developed, and this PCR assay was positive for all the E. faecalis strains tested. Dot blot hybridization using either groES orgroEL as the probe distinguished E. faecalis clearly from other species, indicating that both genes can be used as suitable targets for E. faecalisidentification. Moreover, broad-range PCR-restriction fragment length polymorphism of groESL was designed to differentiate eight commonly encountered Enterococcus species. TheEnterococcus species of reference strains could be easily differentiated on the basis of restriction patterns produced byHaeIII and RsaI. The DNA-based assays developed in this study provide an alternative to currently used methods of identification for clinically important enterococcal species.