Expression of recombinant human tumor suppressor NDPK-A in E. coli
- VernacularTitle:重组人抑癌相关蛋白核苷二磷酸激酶A的原核表达
- Author:
Yanchao RAN
;
Yifei WANG
;
Sheng XIONG
;
Meiying ZHANG
;
Wentao HUANG
;
Linbo LUO
;
Qiuying LIU
- Publication Type:Journal Article
- Keywords:
Nucleoside-diphosphate kinase;
Tumor suppressor proteins
- From:
Chinese Journal of Pathophysiology
1989;0(06):-
- CountryChina
- Language:Chinese
-
Abstract:
AIM: To construct E. coli expression plasmid of recombinant human NDPK-A with a 6?His tag, optimize the expression condition and identify the activity of the product. METHODS: nm23-H1 was subcloned from plasmid pBVNMH1 to pQE40 which contain 6?His purification tag. The expression condition was modulated in grades to get the optimal expression. We purified protein with the Ni+-NTA affinity chromatography column, identified the immunogenicity of the product with Western blot, and measured the kinases activity with HPLC. In addition, angiogenesis inhibition activity of rhNDPK was identified by CAM. RESULTS: The sequence of nm23-H1 subclone in pQE40 was exactly correct. The expression rate of rhNDPK-A was 49 6%. Purified rhNDPK-A specially recognized the antiserum of NDPK-A. It also inhibited angiogenesis. CONCLUSION: PQE-nm23H1 containing 6?His can express target protein at high level. This purification method is simple than other methods, and the product has the same activity as natural human NDPK-A.
