【摘要】 根据细菌的16SrDNA3′端和23SrDNA5′端的高度保守区设计引物,扩增了一株中华鳖Trionyxsinensis"红板病"病原菌嗜水气单胞菌的16S-23SrDNA间区,克隆到pGEM＿T载体上,测序。用BLAST和DNAstar软件对16S-23SrDNA间区序列以及其内tRNA基因与已发表的嗜水气单胞菌和近缘种的IGSG序列进行比较分析,设计出对嗜水气单胞菌特异的PCR引物,建立了检测嗜水气单胞菌的PCR方法,进行了特异性、灵敏性检测,并用此方法检测了不同的水样。更多还原

【Abstract】 Using the conserved regions at 3′ end of 16S and 5′ end of 23S rDNAs as primers, the 16S-23S rDNA IGSs of A. hydrophila, isolated from diseased Chinese turtle (Trionyx sinensis), were amplified and cloned into pGEM-T vector. Different clones were selected to be sequenced and the sequences were analysed with the software BLAST and DNAstar. Based on the results from multiple sequence alignment of representative IGS~G types, a pairs of primers which were specific to A. hydrophila were successfully designed for PCR. Then, a new PCR method was developed and its species-specificity and sensitivity were tested. Finally, this method was used to detect A. hydrophila in different water samples.更多还原