病毒铺膜印迹技术分离牙鲆鳃细胞上的淋巴囊肿病毒结合蛋白

摘要

We used native-PAGE and virus overlay protein binding assay (VOPBA) to identify binding proteins for lymphocystis disease virus in floumder(Paralichthys olivaceus) gill cells(FG). We isolated a 135 kD putative LCDV binding protein molecule. SDS-PAGE and two-dimensional (2-D) gel electrophoresis analysis of this molecule indicated it consisted of three proteins with molecular weights of 58.3 kD, 44.6 kD, and 37.6 kD, of which only the 37.6 kD polypeptide was able to bind to LCDV.%利用非变性电泳与病毒铺膜印迹技术(VOPBA)分离了牙鲆(Paralichthys olivaceus)鳃细胞(FG)上淋巴囊肿病毒结合蛋白,结果显示在FG细胞膜上有分子量为135 kD的蛋白与淋巴囊肿病毒特异结合；对该蛋白切胶回收后进行SDS-PAGE与双向电泳,发现135 kD蛋白由3个蛋白组成,分子量分别为58.3 kD、44.6 kD及37.6 kD; 135 kD蛋白SDS-PAGE的VOPBA显示,仅出现37.6 kD的蛋白带,而58.3 kD、44.6 kD蛋白皆不与淋巴囊肿病毒结合.结果表明牙鲆FG细胞上135kD蛋白是淋巴囊肿病毒的结合蛋白,其37.6 kD蛋白具有病毒结合活性.