人IL-2受体α链cDNA体外转录产物的表达

Expression of human IL-2 receptor chain cDNA in vitro transcripts

重组质粒pJSB13由载体pGEM3多克隆位点中插入人IL-2受体α链cDNA12-937bp片段构成。本文报导了利用载体上的SP6启动子,在体外合成了与插入cDNA片段互补的正链RNA,并用帽子结构类似物在此RNA 5′端进行了加帽,此加帽RNA能在兔网织红细胞体外翻译体系和爪蟾卵母细胞中有效翻译,且翻译产物能分泌到卵母细胞培养液中。

Recombinat plasmid pJSB13 has an insert of 121-937 bp of human IL-2R_α cDNA in multiple cloning site of pGEM3 vector.This plasmid was digested with EcoR I to linearize it and then in vitro transcripted with SP6 RNA polymerase and capped with cap structure analog 7mG (5') G. The capped transcripts were translated in rabbit reticutocyte lysate or Xenopus Iaevis oocytes respectively. The results demonstrated that they can be efficiently translated as functional mRNA in both systems. Moreover, the translation product, IL-2R_α,can be secreted into oocvte culture medium. The program we had used enables one to produce and identify mRNA and conse quently protein from any cDNA clone. Furthermore this research can be of much helpful to the researches on the relationship between structure, function and expression of IL-2R_α.