蓖麻蚕核型多角体病毒包涵体蛋白的提纯及SDS-PAGE分析

THE PURIFICATION AND SDS-POLYACRYLAMIDE GEL ELECTROPHORESIS ASSAY FOR THE POLYHEDRIN IN philosamia cynthia ricini NUCLEAR POLYHEDROSIS VIRUSES

对蓖麻蚕核型多角体病毒 (PhilosamiacynthiariciniNuclearPolyhedrosisViruses)的多角体蛋白 polyhedrin的碱解性质以及分子量等进行了分析。结果发现 :多角体加热灭活和碱解后 ,经等电点沉淀的 polyhedrin通过凝胶电泳仍存在较多的小分子量蛋白带 ,表明通常的 70℃加热不能完全灭活碱性蛋白酶 ;在电泳图谱的主带polyhedrin一侧尚有一伴随的的卫星带 ,可能提示包涵体的外膜蛋白。Sephadex -G2 0 0柱层析纯化的 polyhedrin经SDS -PAGE分析 ,多角体蛋白有 4条带 ,其中主带分子量约为 30KD ,另有一带处于 6 1KD位置 ,推测为 polyhedrin的二聚体 ,说明polyhedrin多聚体在杆状病毒包涵体中的普遍性。

This experiment announced the alkaline degradation property of the polyhedrin in Philosamia cynthia ricini nuclear polyhedrosis viruses and its molecular weight.The result showed that the polyhedra was degradated by alkaline even after heating for inactivation.The polyhedrin was deposited at its isoelectric point.SDS-PAG electrophoresis showed that the polyhedrin was consisted of several polypeptides in low molecular weight,which implied that common 70℃ heating can not inactivate the alkaline protease completely.The satellite band,associated the major polyhedrin,may be the polyhedral envelope protein.After purification by chromatography of Sephadex-G200 column,polyhedrin had 4 polypeptides in SDS-PAGE chart and the molecular weight of the major band was 30KD,the second band was 61KD,which may be the dimer of the polyhedrin.It suggested that the polymer existed generally in the baculovirus polyhedra.