两株芽孢杆菌的鉴定及淀粉酶基因的克隆

摘要

对分离得到的两个菌株(B.H和B.M)进行鉴定,并对它们在芽孢杆菌培养基和淀粉富集培养基上的形态进行观察,通过对其16S rDNA序列的分析,构建系统进化树,确定B.H为枯草芽孢杆菌,B.M为解淀粉芽孢杆菌.根据已报道的枯草芽孢杆菌和解淀粉芽孢杆菌淀粉酶基因保守区域设计引物,分别以两个菌株的基因组DNA为模板,采用PCR扩增的方法成功获得两个淀粉酶基因,其编码区的长度分别为1 434 bp和1 563 bp,为该菌株的进一步利用开发提供了数据支持.%Two isolated strains of Bacillus(B. H and B. M)were identified. Morphologic observation on the Bacillus medium and the starch enrichment medium, and the analysis of the 16S rDNA sequences and Phylogenetic Trees showed that B. H is Bacillus subtilis and B. M is Bacillus amyloliquefaciens. The primes were designed according to conservative sequences of amylase of Bacillus, using each genomic DNA of two strains as templates. The lengths of coding area are 1 434 bp and 1 563 bp respectively. These genes provide the experimental data for further genetic and industrial development of Bacillus.