A cDNA encoding melittin from a wasp (Polistes hebraeus) was amplified and cloned into the GST fusion expression vector pGEX-4T-2. The expressed protein appeared on the SDS-PAGE profiles with an about 29 kDa band. Wes...A cDNA encoding melittin from a wasp (Polistes hebraeus) was amplified and cloned into the GST fusion expression vector pGEX-4T-2. The expressed protein appeared on the SDS-PAGE profiles with an about 29 kDa band. Western blotting proved that the recombinant protein is the fusion protein of GST-PhM. The expression conditions of GST-PhM fusion protein for E. coli BL21 transformant were optimized. Thin layer scanning on the SDS-PAGE profiles showed that the expressed target protein had accumulated up to about 10%～12% of total protein of bacterial cells under the optimized expression condition. Purified and recovered recombinant melittin of Polistes hebraeus showed bioactivity in activating rabbit platelets to aggregate.展开更多
基金Item supported by national natural sciencefoundation of China(No.30271008)Foundation of school ofagriculture and biology of SJTU(AE150054)
文摘A cDNA encoding melittin from a wasp (Polistes hebraeus) was amplified and cloned into the GST fusion expression vector pGEX-4T-2. The expressed protein appeared on the SDS-PAGE profiles with an about 29 kDa band. Western blotting proved that the recombinant protein is the fusion protein of GST-PhM. The expression conditions of GST-PhM fusion protein for E. coli BL21 transformant were optimized. Thin layer scanning on the SDS-PAGE profiles showed that the expressed target protein had accumulated up to about 10%～12% of total protein of bacterial cells under the optimized expression condition. Purified and recovered recombinant melittin of Polistes hebraeus showed bioactivity in activating rabbit platelets to aggregate.
