文献类型：期刊文献

中文题名：3种水产病原弧菌(Vibrio)的16S—23S rDNA间区(IGSs)的克隆、测序与分析

英文题名：CLONING,SEQUENCING AND ANALYSIS OF 16S—23S rDNA INTERGENIC PACERS (IGSs) OF 3 SPECIES OF VIBRIO,THE PATHOGENS IN AQUACULTURE

作者：邓先余[1];王智学[2];孙成波[3];何建国[2]

机构：[1]湖南科技大学生命科学学院;[2]中山大学生命科学学院,广州510275;[3]湛江海洋大学水产学院,湛江524025

年份：2006

卷号：37

期号：2

起止页码：162

中文期刊名：海洋与湖沼

外文期刊名：Oceanologia Et Limnologia Sinica

收录：CSTPCD、、北大核心2004、Scopus、CSCD2011_2012、北大核心、CSCD

基金：国家"863"计划资助项目;2001AA622020号;广东省科技厅专项资助项目;2KB05301N号;湖南科技大学博士基金资助项目;E50437号

语种：中文

中文关键词：溶藻弧菌;河流弧菌;创伤弧菌;16S-23S;rDNA间区;tRNA基因

外文关键词：Vibrio alginolyticus , V. fluvialis , V. vulnificus , 16S-23S rDNA IGS, tRNA gene

中文摘要：根据细菌16S rDNA3端和23S rDNA5端的高度保守区设计引物,PCR扩增了5株溶藻弧菌、2株河流弧菌和2株创伤弧菌的16S—23S rDNA间区,克隆到pGEM-T载体上,测序;采用BLAST和DNAstar软件对16S—23S rDNA间区序列及其内的tRNA基因进行比较分析研究。结果表明,3种弧菌共测出的16S—23S rDNA间区有33条,类型有6种:IGSGLAV、IGSGLV、IGSIA、IGSG、IGSA和IGS0,其中IGSIA和IGSG最为常见,9株弧菌均包含有此两类IGS;在3种弧菌中,大部分IGS序列tRNA基因两端的非编码区具有较高的种内同源性,可以成为设计种特异性引物和探针的靶区。16S—23S rDNA间区结构的差异为建立一类新的弧菌检测方法奠定了基础。溶藻弧菌的16S—23S rDNA间区序列为首次报道。

外文摘要：Based on the bacterial conservative sequences flanking the 3＇ end of the 16S and the 5＇end of the 23S rDNAs, the authors designed primers and PCR-amplified the 16S-23S rDNA intergenic spacers （IGSs） of 5 strains of Vibrio alginolyticus, 2 strains of V.fluvialis and 2 strains of V. vulnificus, all of which are pathogens in aquaculture, and cloned the purified PCR production with pGEM-T vector. Different clones were selected for sequencing. Characteristics of the sequences determined for the 16S-23S rDNA IGS of these 3 species were examined against GenBank using NCBI GenBank BLAST function. Then, the 16S-23S IGS sequences were also specially aligned for intra-species sequence homology analysis using DNAstar software. 33 IGS sequences were deposited in GenBank under accession numbers AY245198 to AY245230, and the 16S- 23S rDNA IGSs of V. alginolyticus were reported for the first time in the world. The results showed that 33 IGSs were sequenced and 6 types of polymorphic IGSs were recognized, namely, IGS^GLAV、IGSGL^GLV、IGS^IA、IGS^G、IGS^A and IGS^O. Among these 6 IGS types, IGS^GLAV is the biggest one, including clusters of tRNA^Glu- tRNA^Lye - tRNA^Ala- tRNA^Val; IGS^GLV , consisting that of tRNA^Glu- tRNA^Lys- tRNA^Val ; IGS^AG, tRNA^Ala- tRNA^Gla ; IGS^IA, tRNA^He- tRNA^Ala ; IGS^G, tRNA^Glu ; IGS^A, tRNA^Ala ; and IGS^O is the smallest one having no tRNA gene. IGS^IA and IGS^G were the most frequent types, both were found in each of these 9 Vibrio strains. Intra-species multiple alignment of all the IGS sequences revealed several highly conserved sequence blocks in non-coding regions flanking the tRNA genes in all of the strains, the first 40 and last 200 nucleotides were most notably, which can be targeted to design species-specific PCR primers or detection probes. The structural variation of the spacers would help develop diagnostic methods for these pathogenic Vibrios.