米曲霉giF-10内切葡聚糖酶基因在大肠杆菌中的表达

【目的】克隆米曲霉giF-10内切葡聚糖酶基因并检测其在大肠杆菌中的表达。【方法】以自行分离筛选出的天然米曲霉giF-10的基因组DNA为模板,PCR高保真扩增,扩增产物连接到pMD19-T克隆载体上,并构建原核表达载体,转化大肠杆菌BL21(DE3)。【结果】序列分析表明,其长度为1 251bp,编码417个氨基酸,序列比对发现与内切葡聚糖酶基因CelB序列相似性高达100%,将此基因注册GenBank(HQ739052)。刚果红水解圈筛选结果表明已成功表达。【结论】克隆了米曲霉giF-10内切葡聚糖酶基因,并在大肠杆菌成功表达,为纤维素酶工业化生产奠定基础。

Expression of Aspergillus oryzae giF-10 Endoglucanase Gene in Escherichia coli

【Objective】 The aim of this research was to clone the gene encoding endoglucanase from Aspergillus oryzae giF-10 and detect the expression of the gene in Escherichia coli BL21(DE3).【Method】 The gene encoding endoglucanase was cloned by PCR from Aspergillus oryzae giF-10,and inserted into the expression vector pET32a to express by IPTG induction in Escherichia coli.【Results】 The gene was 1251 bp,encoding 417 amino acids,and was registered in GenBank(Accession number HQ739052).It shared a 100% sequence identity with the endoglucanase CelB gene by sequence alignment.The expression construct was transferred into Escherichia coli BL21(DE3) and expressed successfully through hydrolysis cycle test.【Conclusion】 The gene encoding endoglucanase was cloned from Aspergillus oryzae giF-10 and expressed successfully in Escherichia coli BL21(DE3),which is important for industrial production of cellulase.