摘要
体外培养后分离出的3个伊氏锥虫变异群体,经环磷酰胺处理大鼠(CY 大鼠)扩增收虫。虫体纯化后超声波裂解,超速离心制成可溶性抗原,经 Sephadex G-50脱盐和 DE-52离子交换层析提纯可变表面糖蛋白(VSG)。经 SDS-PAGE 鉴定,所提纯的3个 VSG 分子量均为40500。用日立835-50型高速氨基酸组成分析仪分析,3个 VSG 均由 Asp,Pro,Thr,Ser,Glu,Gly,Ala,Cys,Val,Met,Ile,Leu,Tyr,Phe,Lys,His,Arg共17种氨基酸组成,但在组成比例上差异明显,尤其表现在 Pro,Met,Leu,Ile.Tyr 和 Clu。经 Western blot-ting 鉴定,提纯的3种 VSG 是3种不同的特异性抗原。
Three VAT-derived populations were expanded in cyclophosphamide treated(CY)rats for experimen- tal use.Purified trypanosomes were broken by high-speed homogenization and centrifugated by high g to form soluble proteins.The soluble proteins were desalted on a column of Sephadex G-50 prior to chromatography on a DE-52 column.The material in the first peak constituted the purified VSG.The molecular weight of the purified VSG was 40 500 by detection of SDS-PAGE.The differences of three VSGs in molecular weight were small.Three VSGs were composed of 17 amino acids by analysis of Model 835-50 high speed amino acid analyzer.They were Asp,Pro,Thr, Ser,Glu,Gly,Ala,Cys,Val,Met,He,Leu,Tyr,Phe,Lys,His,and Arg.The differences in the proportion of amino acid were most striking especially in Pro,Mei,Leu,He,Tyr,and Glu.Three purified VSGs were identified as specific antigens of three variant coats by Western blotting.
出处
《南京农业大学学报》
CAS
CSCD
北大核心
1996年第1期68-72,共5页
Journal of Nanjing Agricultural University
基金
国家教委博士点专项科研基金
江苏省自然科学基金
关键词
伊氏锥虫
可变表面糖蛋白
抗原变异
Trypanosoma evansi
variable surface glycoprotein
antigenic variation