摘要
OaEPV经2×SDS包涵体碱性裂解缓冲液裂解,离心,分别用上清液及沉淀与绿僵菌孢子混合感染黄胫小车蝗,上清液的增效活性比沉淀大2.5倍。用三种不同方法(2×SDS包涵体碱性裂解液法,0.02 mol/L NaOH裂解法,8 mol/L尿素裂解法)裂解OaEPV所得蛋白液与绿僵菌混用,生测结果表明,包涵体裂解液法制备所得蛋白增效活性最高,NaOH裂解法次之,尿素裂解法制备的蛋白几乎没有增效活性。将OaEPV用包涵体碱性裂解液裂解后,用葡聚糖G-200凝胶柱层析进行分离,出现两个洗脱峰,这两个峰的洗脱液浓缩后进行生物测定,初步表明增效作用主要为第二个峰的蛋白片段。经SDS-聚丙烯酰胺凝胶电泳,第二个峰的蛋白片段的分子量为40kDa左右。
Oedaleus asiaticus entomopoxvirus(DaEPV) contains a factor which enhances the infection of Metarhizium flaroviride in the grasshopper Oedaleus infernalis. When the spheroids of OaEPV were dissolved with 2×SDS spheroid alkaline solution and centrifuged, the enhancing activity of the supernatant was 2.5 times higher than in the pellet. The enhancing activity of spheroid proteins depended on the alkali used to dissolve the spheroids. The greatest activity was detected with spheroid alkaline solution, followed with 0.02 mol/L NaOH, and none with 8 mol/L urea. Spheroid proteins were fractionated on a sephadex G-200 column. Two fractions were obtained and the second fraction had the enhancing activity. SDS-PAGE indicated the molecular weight of the active fraction was 40 kDa.
出处
《中国病毒学》
CSCD
2003年第5期478-481,共4页
Virologica Sinica
基金
国家自然科学基金(39770512)
