纤维素降解菌N2-10菌株β-1,4-内切葡聚糖酶基因的克隆及表达

Cloning and expression of β-1,4-endoglucanase gene of cellulose derading bacteria N2-10

为探究枯草芽孢杆菌(Bacillus subtilis)N2-10菌株降解纤维素的机理,通过同源性比对设计兼并引物扩增菌株β-1,4-内切酶葡聚糖基因,并进行生物信息学分析,然后研究其在大肠杆菌中的表达情况.结果显示,该内切葡聚糖酶基因大小为1500bp,共编码499个氨基酸,具有典型的纤维素酶结构,SDSPAGE电泳检测其表观分子质量约为55ku;刚果红染色显示,表达产物具有纤维素酶活性.

To validate the mechanism of cellulose degradation by Bacillus subtilis N2-10,through homology comparison,the degenerate primers was designed and theβ-1,4-endoglucanase gene of B.subtilis N2-10 was amplified.By useing bioinformatics tools in analyzing the sequences of genes and proteins and considering that the expression of theβ-1,4-endoglucanase gene in E.coli,the result shows that,this gene is 1 500 bp,which could totally encodle 499 amino acids.This study also found that it had the typical structure of cellulose.SDS PAGE analysis found that the apparent molecular weight of the expression product was about 55 ku.Congo red staining analysis found that the expression product had the activity of cellulose.