Expression of recombinant human tumor suppressor NDPK-A in E. coli |
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| Authors: | RAN Yan-chao WANG Yi-fei XIONG Sheng ZHANG Mei-ying HUANG Wen-tao LUO Lin-bo LIU Qiu-ying |
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| Institution: | Biomedicine Research and Development Center, Jinan University, Guangzhou 510632, China |
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| 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. |
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| Keywords: | Nucleoside-diphosphate kinase Tumor suppressor proteins |
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