摘要
以细菌的 1 6SrDNA 3′端和 2 3SrDNA 5′端的高度保守区为引物 ,扩增了 3株杂色鲍致病菌—副溶血弧菌(Vibrioparahaemolyticus)的 1 6S 2 3SrDNA间区 ,克隆到pGEM T载体上 ,测序。用BLAST和DNAstar软件对 1 6S 2 3SrDNA间区序列及其内的tRNA基因进行比较分析。结果表明副溶血弧菌ZSU0 0 8和ZSU0 0 9测出的 1 6S 2 3SrDNA间区均有 6条 ,间区类型也相同 ,分别为IGSGLAV 、IGSGLV、IGSAG、IGSIA、IGSG 和IGS0 。其中IGSGLAV最大 ,包含tRNAGlu、tR NALys、tRNAAla和tRNAVal基因 ;IGSGLV包含tRNAGlu、tRNALys和tRNAVal基因 ;IGSAG包含tRNAAla和tRNAGlu基因 ;IGSIA,则为tRNAIle和tRNAAla基因 ;IGSG 仅包含tRNAGlu基因 ;而IGS0 最小 ,未包含任何tRNA。菌株ZSU0 1 0测出的 1 6S 2 3SrDNAIGS序列有 5条 ,除缺少IGSAG 外 ,其余的IGS类型均与ZSU0 0 8和ZSU0 0 9相同。与GenBank内的副溶血弧菌IGS序列比较 ,发现副溶血弧菌所有类型的IGS的tRNA基因两端的非编码区具有较高的种内同源性。 1 6S 2 3SrDNA间区结构的差异为建立一种新的副溶血弧菌检测方法奠定了基础。
Using the conserved sequences flanking the 3′ end of the 16s and the 5′ end of the 23S rDNAs as PCR primers, the 16S-23S rDNA intergenic spacers (IGSs) of 3 strains of Vibrio parahaemolyticus, which is a pathogen of abalone(Haliotis diversicolor), were amplified and cloned with pGEM-T vector. Different clones were selected to be sequenced and the sequences were analysed with the software BLAST and DNAstar. Analysis of the IGS sequences suggested that both of the strain ZSU008 and ZSU009 contain six types of polymorphic 16S-23S spacers, namely, IGS GLAV, IGS GLV, IGS AG, IGS IA, IGS G and IGS 0; but the strain ZSU010 lacks IGS AG. Among these six IGS types, IGS GLAV is the biggest type, including the cluster of tRNA Glu-tRNA Lys-tRNA Ala-tRNA Val; IGS GLV, including that of tRNA Glu-tRNA Lys-tRNA Val; IGS AG, tRNA Ala-tRNA Glu; IGS 0 is the smallest, which has no tRNA gene. Multiple alignment of all the IGS sequences of these 3 strains with those of V. parahaemolyticus IFO12711 available from GenBank revealed some regions of similarity in the non-coding regions flanking the tRNA genes within all the IGS types. The structure variation of the spacers lays a foundation for developing a diagnostic method for V. parahaemolyticus.
出处
《微生物学报》
CAS
CSCD
北大核心
2004年第3期304-308,共5页
Acta Microbiologica Sinica
基金
国家"8 63计划"( 2 0 0 1AA62 2 0 2 0 )
广东省科技厅专项资助项目 ( 2KB0 5 3 0 1N)~~
