西方许旺酵母α-淀粉酶基因的克隆及其在大肠杆菌中的分泌表达

Cloning Expression α-Amylase Gene from Schwanniomyces occidentalis

目的:将带有完整自身信号肽的西方许旺酵母α-淀粉酶基因克隆到大肠杆菌中,验证西方许旺酵母α-淀粉酶基因能否在大肠杆菌中有效表达。方法:利用PCR扩增带有完整自身信号肽的西方许旺酵母α-淀粉酶基因,并将其接入Zeocin启动子片段,构建了重组表达载体GapZA,转化大肠杆菌,验证得到的阳性克隆菌株是否表达α-淀粉酶活性。结果:阳性克隆菌株均有α-淀粉酶活性。结论:证明了许旺酵母α-淀粉酶能在自身信号肽引导下分泌到大肠杆菌细胞外,并且表现出明显酶活。

Objectives： The α- Amylase was amplified from Schwanniomyces occidentalis using PCR Technique.The amplified 1.65kb DNA fragment was inserted into the vector GapZA, which was cloned in E. coli BL2. The transter can secrete the α- Amylase into the medium. The rnolecular weight of the α - Amylase was about 56kD by SDS - PAGE.