Vero毒素-1的纯化及特性分析  

Purification and Characterization of Vero toxin 1

作　　者：李海波[1] 王棣[1] 赵红[1] 孟筱琦[1] 

机构地区：[1]兰州生物制品研究所,兰州730046

出　　处：《微生物学免疫学进展》2001年第1期11-14,共4页Progress In Microbiology and Immunology

摘　　要：从含有VT1全基因的基因工程菌中纯化出VT1。纯化的步骤包括 (NH4 ) 2 SO4 盐析 ,两次DEAESepharoseFastFlow柱层析。最终从 4L培养物中纯化出 1 5mg纯毒素 ,收率为 8 6 % ,梯度Native PAGE测定毒素的分子量为 70kD ,SDS PAGE电泳表明毒素有两个亚基 ,分子量分别是 32kD和 7 7kD。对VT1的多种生物学特性进行了研究 ;经测定VT1对Vero细胞的半数致死量CD50 为 1pg ,对小鼠的半数致死量LD50 为 18ng ,引起兔肠襻积液的最小毒素量是 1 2 5Vero toxin 1 was purified to homogeneity from Escherichia coli MC1061 which contains the toxin genes cloned in the multicopy plasmid pUC118.The scheme involved(NH 4) 2SO 4 precipitation and repeated anion exchange chromatography.About 1.5 mg purified toxin was obtained from cell extracts of 4 liters of culture with a yield of about 8.6%.Gradient Native PAGE(4%-30%)showed the molecular weight of the toxin to be 70 000 and SDS PAGE showed that the toxin consisted of A and B subunits with apparent molecular weights of 32 000 and 7 700 respectively.The purified vero toxin 1 showed various biological activities such as cytotoxicity to vero cells and killing about 50% of cells at 1 pg,lethal toxicity to mice by injected intraperitoneally,the LD 50 is 18 ng,the fluid accumulation in rabbit ileal loops at concentration of 1.25 μg/loop.

关 键 词：VERO毒素-1 纯化 特性 大肠杆菌 

分 类 号：R378[医药卫生—病原生物学]