关键词: 鸡胚, 组织膜蛋白, 鸡毒霉形体黏附蛋白, Dot-ELISA

Abstract: The GST-pMGA was expressed in E. coli BL21 with the pGEX fusion expression system. GST-pMGA expressed in supernatant was purified using GST Bind Resin and a plenty of soluble protein was obtained, the final preparation was 96% after purification by SDS-PAGE analysis. GST-pMGA was incubated with thrombin protease overnight, the pMGA was obtained and showed well immunological activity by Western-blot analysis. The tissues membrane protein from trachea, heart, lung, liver, kidney, thymus, bursa of Fabricius, spleen, proventriculus and the duodenum of SPF chick embryo were obtained through centrifugation, Dot-ELISA was applied to detect the specific binding relation between the membrane protein and pMGA. The results showed that pMGA and the tissues membrane protein of chick embryo had a specific binding except the spleen, this binding indicated receptor of pMGA on the tissues membrane protein of chick embryo. In membrane protein extracted from these tissues except spleen, there are the same product with Molecular weight approximately 30 kD by SDSPAGE analysis, so it was concluded it’s the possible protein interacting with pMGA. All this makes it possible to do further studies of receptor of pMGA.

Key words: chick embryo, the tissues membrane protein, pMGA, Dot-ELISA