芽胞杆菌β-甘露聚糖酶基因部分序列的克隆及相似性分析

Cloning and Alignment of the Partial Mannanase Gene of Bacillus spp

摘要:

通过平板筛选,从28株芽孢杆菌中筛选出16株产β-甘露聚糖酶的菌株。利用一对兼并引物,通过PCR分别从不同来源芽孢杆菌基因组DNA上扩增出β-甘露聚糖酶编码基因的一段保守序列。将基因片段测序并同已发表的β-甘露聚糖酶基因进行相似性分析,并构建进化树。结果表明:克隆到的β-甘露聚糖酶部分基因序列与报道的相比,其最高同源性在62%～98%之间。环形芽孢杆菌β-甘露聚糖酶编码基因间相似性较低,枯草芽孢杆菌及其它芽孢杆菌的β-甘露聚糖酶编码基因间相似性较高。

Abstract:

By functional plates,16 strains which can produceβ-mannana-se were isolated frnm 28 Bacillus spp.Using a pair of degenerated primers,the conserved fragments ofβ-mannanase gene from the selected strains were amplified by PCR.The obtained nucleotide fragments were sequenced and compared with the homologousβ-mannanase genes in GenBank and a phylogenetic tree was generated.Comparing to the genes codingβ-mannanase published,the cloned nucleotide fragments show the highest sequence identity between 62% and 98%.The genes coding for β-mannanase of Bacillus circulus have low identity while the β-mannanase genes of Bacillus subtilis and other Bacillus spp. have high identity.