纳豆激酶原的基因克隆及在大肠杆菌中的表达

目的　构建纳豆激酶原基因克隆 ,实现其在大肠杆菌中的表达。方法　从纳豆芽胞杆菌中分离纯化基因组DNA ,作为模板通过PCR扩增纳豆激酶原基因。运用重组DNA技术 ,构建表达质粒pESX 1,转化大肠杆菌JF112 5 ,温度诱导表达目的蛋白。结果　琼脂糖凝胶电脉及核苷酸序列分析证实 pESX 1为含纳豆激酶原基因的阳性克隆。在大肠杆菌中表达纳豆激酶原 ,经自体加工产生具有纤溶活性的纳豆激酶。结论　成功地构建了纳豆激酶原基因克隆 ,实现了在大肠杆菌中的表达。

Molecular Cloning and Expression of Pro nattokinase Gene in Escherichia coli

Purpose Molecular cloning and expression of pro nattokinase gene with E.coli . Methods Chromosome DNA was isolated from Bacillus natto .The pro nattokinase gene was then amplified from chromosome by PCR.The expression vector pESX?1 was constructed and transformed into E.coli JF1125. Results The agarose gel electrophoresis and nucleotide sequencing confirmed that pESX?1 was the positive clone.Pro nattokinase was expressed and then produced active nattokinase through an intramolecular self processing mechanism. Results The agarose gel electrophoresis and nucleotide sequencing confirmed that pESX?1 was the positive clone.Pro nattokinase was expressed and then produced active nattokinase through an intramolecular self processing mechanism. Conclusions A clone encoding pro nattokinase was constructed and pro nattokinase gene was expressed with E.coli .