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《Journal of Wuhan Botanical Research》 2008-01
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Cloning and Prokaryotic Expression of Alpha-momorcharin Gene

OUYANG Yong-Chang1,2,3,ZHUANG Dong-Hong1,HU Zhong1,LI Xiang2(1.Department of Biology of Shantou University,Shantou,Guangdong 515063,China;2.South China Sea Institute of Oceanology,The Chinese Academy of Sciences,Guangzhou 510301,China;3.The Graduate School of Chinese Academy of Sciences,Beijing 100039,China)  
Momordicin(Alpha-momorcharin)is a kind of multifunctional ribosome inactivating protein(RIP)with anti-tumor,anti-bacterial,anti-HIV and immunosuppressive activity.In this study,the gene encoding mature Alpha-momorcharin protein was amplified from the genomic DNA of Momordica charantia L.by using PCR(polymerase chain reaction)method.The amplified product was cloned into expression vector pET28a(+),and then expressed in the BL21(DE3)and RosettaTM(DE3)pLysS.Only in the RosettaTM(DE3)pLysS strain,the 33-kD-weight recombinant protein of Alpha-momorcharin was successfully expressed and the soluble recombinant protein was occurred when the strain was induced with IPTG at 18℃ and 22℃ temperature.The soluble recombinant protein was purified by the Ni-NTA resin and followed by Western-blotting analysis.This result revealed that the purified recombinant protein could be recognized by His-Tag antibody.It lays a foundation for further studies of the function and activity of the Alpha-momorcharin recombinant protein.
【Fund】: 广东省高校自然科学研究项目(Z02036)~~
【CateGory Index】: Q943.2
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