溶藻弧菌中类霍乱弧菌毒力岛转座酶基因及侧翼序列分子生物学特征分析  

Analysis of molecular biological characteristic of the gene and its flanking sequences,similar with transposase in Vibrio cholerae pathogenicity island,among V.alginolyticus strains

作　　者：罗鹏[1] 胡超群[1] 

机构地区：[1]中国科学院南海海洋研究所,中国科学院海洋生物资源可持续利用重点实验室,广州510301

出　　处：《微生物学报》2008年第10期1367-1372,共6页Acta Microbiologica Sinica

基　　金：国家“973项目”(2006CB101803);国家自然科学基金(30700016)~~

摘　　要：【目的】调查类似霍乱弧菌毒力岛(VPI)转座酶(vpiT)的基因是否在溶藻弧菌中分布,并了解其全序列及侧翼序列的分子生物学特征。【方法】对94株溶藻弧菌是否携带类似VPI的vpiT基因进行PCR检测,对阳性株进行了PCR产物直接测序,根据获得的部分已知序列,设计引物,通过反向PCR扩增出全长类似vpiT的基因valT及部分侧翼片段,对反向PCR产物进行克隆测序,然后对获得的valT及侧翼序列进行生物信息学分析。【结果】发现94株溶藻弧菌中只有从粤东对虾池水分离的2个株E06011、E0612在PCR检测中产生了预期扩增片段。测序表明两者序列(valT-S1)完全一致。根据反向PCR及克隆最终获得的溶藻弧菌E0601全长valT基因及部分侧翼序列valT-S3。对valT-S3生物信息学分析表明:valT是一个高度类似于霍乱弧菌毒力岛vpiT的转座酶基因。【结论】根据上述结果及相关文献,有理由相信valT基因及其侧翼片段是异源获得,霍乱弧菌VPI元件或整体很可能在包括溶藻弧菌在内的弧菌种间转移。[Objective] To investigate whether the gene similar with transposase gene （vpiT） from pathogenicity island of Vibrio cholerae exists in V alginolyticus strains, and to analyze molecular biological characteristic of the gene and its flanking sequences. [Methods] PCR detection of the gene, similar with vpiT in pathogenicity island of V. cholerae was done among 94 strains of V. alginolyticus,. PCR products from positive strains were directly sequenced. Based on acquired partial sequences, we designed primers for reverse PCR, and got the amplification fragment containing complete gene （vaIT） from V. alginolyticus E0601, which was similar with vpiT gene. The reverse PCR product was cloned and sequenced, and the acquired sequence was analyzed with bioinformatic methods. [Results] We found that among 94 V. alginolyticus strains, only V. alginolyticus E0601 and E0612, from east coastal areas of Guangdong province, produced predicted positive amplification fragments in PCR detection. Sequencing indicated that amplification fragments from V. alginolyticus E0601 and E0612 had identical DNA sequence （named valT-S 1）. Sequence valT-S3 from V alginolyticus E0601, containing complete valT gene and flanking segments, was finally obtained through reverse PCR, cloning, and sequencing. Bioinformatic analysis on valT-S3 suggested that valT was transposase gene, highly similar with vpiT in V. cholerae VPI. [Conclusion] According to above result and related references, we believe that valT and its flanking segments were acquired from heterogenous bacteria, and VPI or its component probably transfers among Vibrio species including V alginolyticus.

关 键 词：溶藻弧菌 霍乱弧菌 毒力岛 转座酶 

分 类 号：Q93[生物学—微生物学]