Keywords

¦¡- Momourin, Rosettatm (De3) Plyss Strain, Recombinant Protein

Abstract

Alpha-momorcharin is a multifunctional ribosome-inactivating protein with anti-tumor, anti-HIV-1 and antibacterial activities, as well as immunosuppressive activity. The mature ¦Á-moumourin protein gene was amplified from the genome of bitter melon using PCR (polymerase chain reaction) technology, and was subcloned into the expression vector pet28a(+), and then carried out in bl21(de3) and rosettatm(de3)plyss respectively. Express. The results show that only in rosettatm(de3)plyss, the recombinant ¦Á-moumourin gene can successfully express a 33kd recombinant protein, and at temperatures of 18¡ãC and 22¡ãC, after induction by iptg, a large amount of Soluble recombinant protein. Using the 6¡Áhis tag on the N terminus of the recombinant protein, the ¦Á-moumourin recombinant protein was purified through ni-nta column. Western hybridization experiments showed that the purified recombinant protein could react specifically with mouse anti-his-tag monoclonal antibodies. The acquisition of the recombinant protein of ¦Á-moumourin laid the foundation for further systematic research and modification of its function and activity.

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Original research done by Ouyang Yongchang, Zhuang Donghong, Hu Zhong, Li Xiang

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