摘要
采用ｐＥｘＳｅｃⅠ载体系统进行了蓖麻毒素Ａ链的原核表达，经ＣＭ－Ｓｅｐｈａｒｏｓｅ一步纯化后，获得了纯度约８０％的重组蓖麻毒素Ａ链．将其与几种天然单链核糖体失活蛋白进行了超螺旋ＤＮＡ裂解研究和无细胞体系中蛋白合成抑制试验，结果表明，重组蓖麻毒素Ａ链具有类似于天然单链核糖体失活蛋白的活性。
Recombinant ricin toxin A chain(rRTA)is one of the important elements used for constructing immunotoxin,which may be useful as a treatment of many diseases.To obtain rRTA,ricin toxin A chain gene was cloned into pExSecⅠ vector and expressed in E.coli BL21(DE3),the proportion of expressed rRTA in total bacteria protein was about 20%.The expression products obtained a purity of 80% or so by a CM Sepharose column.Recombinant ricin toxin A chain,and several wild type single chain ribosome inactivating proteins,were analyzed by cleavage assay of supercoiled DNA and 3 H Leu labeled protein synthesis inhibition assay.The results suggest that rRTA had the similar activity as the others while there had no linear correlation between the two methods.
出处
《中国生物化学与分子生物学报》
CAS
CSCD
1999年第1期71-74,共4页
Chinese Journal of Biochemistry and Molecular Biology
基金
国家"863"生物技术项目
关键词
蓖麻毒素A链
核糖体失活蛋白
无细胞体系
Recombinant ricin toxin A chain
Ribosome inactivating proteins
Cell free system
