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

机译:化学合成的人类免疫干扰素基因在大肠杆菌中的表达。

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摘要

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.
机译:完全合成了由人免疫干扰素(hIFN-γ)基因,起始和终止信号以及适当的限制性核酸内切酶位点组成的双链DNA的454个碱基对片段。合成涉及通过快速,固相程序和寡核苷酸的酶促连接制备62个寡脱氧核糖核苷酸。该合成基因在lac UV5启动子的控制下在大肠杆菌中表达。该产物具有抗病毒活性,该活性对酸不稳定,并且被针对hIFN-γ的抗血清完全中和,但是未被针对hIFN-α或hIFN-β的抗血清完全中和。通过凝胶过滤和SDS-聚丙烯酰胺凝胶电泳,估计大肠杆菌产生的hIFN-γ的分子量分别为约32,000和17,000。

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