Rodgers R E, Chang D, Cai X, Consigli R A
Division of Biology, Kansas State University, Manhattan 66506.
J Virol. 1994 May;68(5):3386-90. doi: 10.1128/JVI.68.5.3386-3390.1994.
A recombinant system for the major capsid VP1 protein of budgerigar fledgling disease virus has been established. The VP1 gene was inserted into a truncated form of the pFlag-1 vector and expressed in Escherichia coli. The budgerigar fledgling disease virus VP1 protein was purified to near homogeneity by immunoaffinity chromatography. Fractions containing highly purified VP1 were pooled and found to constitute 3.3% of the original E. coli-expressed VP1 protein. Electron microscopy revealed that the VP1 protein was isolated as pentameric capsomeres. Electron microscopy also revealed that capsid-like particles were formed in vitro from purified VP1 capsomeres with the addition of Ca2+ ions and the removal of chelating and reducing agents.
已建立一种用于虎皮鹦鹉幼雏病病毒主要衣壳VP1蛋白的重组系统。将VP1基因插入截短形式的pFlag-1载体中，并在大肠杆菌中表达。通过免疫亲和层析将虎皮鹦鹉幼雏病病毒VP1蛋白纯化至接近均一。合并含有高度纯化VP1的级分，发现其占原始大肠杆菌表达的VP1蛋白的3.3%。电子显微镜显示，VP1蛋白以五聚体壳粒形式分离出来。电子显微镜还显示，在体外，通过添加Ca2+离子并去除螯合剂和还原剂，由纯化的VP1壳粒形成了衣壳样颗粒。
