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人免疫干扰素化学合成基因在大肠杆菌中的表达

Expression in Escherichia coli of chemically synthesized gene for the human immune interferon.

作者信息

Tanaka S, Oshima T, Ohsuye K, Ono T, Mizono A, Ueno A, Nakazato H, Tsujimoto M, Higashi N, Noguchi T

出版信息

Nucleic Acids Res. 1983 Mar 25;11(6):1707-23. doi: 10.1093/nar/11.6.1707.

Abstract

A 454 base pair fragment of double stranded DNA consisting of a gene for a human immune interferon (hIFN-gamma), initiation and termination signals plus appropriate restriction endonuclease sites, was totally synthesized. The synthesis involved preparation of 62 oligodeoxyribonucleotides by rapid, solid phase procedures, and enzymatic ligation of the oligonucleotides. This synthetic gene was expressed in E. coli under the control of the lac UV5 promoter. The product has antiviral activity which was acid labile and completely neutralized by antiserum to hIFN-gamma but not by antiserum to hIFN-alpha or hIFN-beta. Molecular weight of hIFN-gamma produced by E. coli was estimated to be about 32,000 and 17,000 by gel filtration and SDS-polyacrylamide gel electrophoresis respectively.

摘要

一段由人类免疫干扰素(hIFN-γ)基因、起始和终止信号以及合适的限制性内切酶位点组成的454个碱基对的双链DNA片段被完全合成。合成过程包括通过快速固相程序制备62个寡脱氧核糖核苷酸,并对这些寡核苷酸进行酶促连接。这个合成基因在lac UV5启动子的控制下在大肠杆菌中表达。产物具有抗病毒活性,该活性对酸不稳定,能被抗hIFN-γ血清完全中和,但不能被抗hIFN-α或抗hIFN-β血清中和。通过凝胶过滤和SDS-聚丙烯酰胺凝胶电泳分别估计,大肠杆菌产生的hIFN-γ的分子量约为32,000和17,000。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/feaa/325830/c342baa1871d/nar00351-0088-a.jpg

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