含PR启动子的原核高效表达载体的构建及应用
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国家“863”(九·五)计划资助项目(1030104).


Construction and Application of A High Level Expression Vector Containing PR Promoter
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    摘要:

    组建了一个仅含PR启动子的原核高效表达载体pRC,它同时含有cⅠ调控基因、多酶切位点和两个强的转录终止序列.现已成功地用于表达重组人肿瘤坏死因子-α(hTNF-α)、重组人白细胞介素-3(hIL-3)和抗溶菌酶(HEL)抗体Fd基因,表达量均占菌体总蛋白的36%以上.同时还研究了不同的宿主菌和原核增强子序列等因素对PR启动子载体表达的影响.此外,还比较了分别以PR、PL或PRPL作为启动子时表达hTNF-α的情况,结果表明,单用PR或PL启动子可获得与使用PRPL串联启动子一样的高效表达.

    Abstract:

    A high level expression vector pRC has been constructed, which contains PR promoter, cIts857 gene, multiple cloning sites(MCS) and two strong transcription terminators. With this vector, hTNF-α, hIL-3 and anti-HEL McAb Fd genes have been expressed successfully, and their products each account for more than 36% of the total cell proteins. At the same time, it has been studied whether such factors as procaryon enhancer sequence and different strains affect the expression level of PR promoter vector. Furthermore, the inductive efficiency was compared among PR, PL and PRPL promoter, and the result has showed that PR or PL is as strong as PRPL.

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陈建军,孙苗,陈常庆,卢芳,王德宝.含PR启动子的原核高效表达载体的构建及应用[J].生物化学与生物物理进展,1999,26(5):495-499

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  • 收稿日期:1998-08-20
  • 最后修改日期:1998-12-23
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