一株产淀粉酶枯草芽孢杆菌16S rDNA的克隆及序列分析

16S rDNA sequence cloning and analysis of a high amylaseproduction Bacillus subtilis strain

对实验室保存的一株产α-淀粉酶的枯草芽孢杆菌的16S rDNA区进行克隆及序列分析.采用PCR克隆方法,对其菌株的16S rDNA区进行序列扩增,扩增产物经琼脂糖凝胶电泳,获得一个大小约为1 300 bp的特异性扩增条带,随后将测序结果用GeneBank数据库中的BLAST软件与获取的已知枯草芽孢杆菌的16S rDNA序列进行序列比对分析.结果表明:该枯草芽孢杆菌与枯草芽孢杆菌FZB42具有相似的序列,相似性为97%.根据上述分析结果可判定2种枯草芽孢杆菌为同属细菌.

This experiment adopted the genome which extracted form Bacillus subtilis strain that can hydrolyze amylase as a template,used the PCR cloning method to amplify the 16S rDNA Regions sequence of it.Through the agarose gel electrophoresis of the PCR products,a specific amplification bands in the size of approximately 1 300 bp was got.Used the sequence determination and the comparative analysis of the known Bacillus subtilis’s 16S rDNA sequence which got from the International Molecular Biology Database in Internet,the results showed that,according to the results of the 16S rDNA sequence analysis,it would be found that the similarity between the two strains was about 97%.It can be initially identified that these two strains belong to the same kind.