CAJ | 학술논문

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BL21进行蛋白高表达,经Westem blot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3 mg/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.
목적:구건재대장간균중표체중조인신경형일양화담합매적고표체계통.방법:용PCR방법종cD-NA중확증목적편마기인,연후장편마기인련접입표체재체pCWori+,장중조적질립전염입대장간균BL21진행단백고표체,경Westem blot학인표체후,진행대량배양,통과층석방법정제차매,병용흡수광보법대매적성질진행측정.결과:종해표체계통중가이획득3 mg/L배양기적고산량적일양화담합매.결론:종해표체계통중가획득대량유활성적인일양화담합매.
AIM:To construct a high-level expression system of recombinant human neuronal nitric oxide synthase(hnNOS)full-length enzyme in Escherichia coli.METHODS:The coding sequence of hnNOS full-length was firstly amplified by PCR,and then ligated into the expression vector pCWori + .The recombinant plasmid was transformed into Escherichia coli BL21 for high-level expression.After having been checked with Western blot,the enzyme was used for large-scale culture and purification.Finally,the property of the enzyme was determined by spectrophotometric method.RESULTS:The constructed expression system could give a yielding of 3 mg/L initial culture.CONCLUSION:The expression system constructed is fully sufficient to express the active human neuronal nitric oxide synthase.