论文部分内容阅读
目的 :构建在大肠杆菌中表达重组人神经型一氧化氮合酶的高表达系统。方法 :用PCR方法从cD NA中扩增目的编码基因 ,然后将编码基因连接入表达载体pCWori+,将重组的质粒转染入大肠杆菌BL2 1进行蛋白高表达 ,经Westernblot确认表达后 ,进行大量培养 ,通过层析方法精制此酶 ,并用吸收光谱法对酶的性质进行测定。结果 :从该表达系统中可以获得 3mg/L培养基的高产量的一氧化氮合酶。结论 :从该表达系统中可获得大量有活性的人一氧化氮合酶。
Objective: To construct a high expression system of recombinant human neuronal nitric oxide synthase in Escherichia coli. Methods: The target gene was amplified by PCR from cD NA, then the gene was inserted into expression vector pCWori +. The recombinant plasmid was transfected into E. coli BL21 for high protein expression. After being confirmed by Western blot, a large number of cells were cultured The enzyme was purified by chromatography and its enzymatic properties determined by absorption spectroscopy. Results: A high yield of nitric oxide synthase at 3 mg / L medium was obtained from this expression system. Conclusion: A large number of active human nitric oxide synthase can be obtained from this expression system.