Construction of human neuronal nitric oxide synthase expression system in Escherichia coli
- VernacularTitle:人类神经型一氧化氮合酶原核表达载体的构建和在大肠杆菌中的表达(英文)
- Author:
Jie FU
;
Yubing ZHOU
;
Yingshe ZHAO
;
Zhiwen GUAN
;
Iyanagi TAKASHI
- Publication Type:Journal Article
- Keywords:
Nitric-oxide synthase;
Escherichia coli;
Gene expression
- From:
Chinese Journal of Pathophysiology
1989;0(06):-
- CountryChina
- Language:Chinese
-
Abstract:
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.
