杂交白细胞干扰素的羧基末端区域影响抗病毒特异性。

Carboxyterminal region of hybrid leukocyte interferons affects antiviral specificity.

作者信息

Franke A E, Shepard H M, Houck C M, Leung D W, Goeddel D V, Lawn R M

出版信息

DNA. 1982;1(3):223-30. doi: 10.1089/dna.1.1982.1.223.

DOI:10.1089/dna.1.1982.1.223

Abstract

Four hybrid human leukocyte interferon (IFN-alpha) genes have been constructed and expressed in Escherichia coli using molecular cloning methods. Plasmids containing genes encoding human interferons, IFN-alpha A, IFN-alpha D, IFN-alpha I and several hybrids of the different IFN-alpha genes (formed by in vitro recombination at common restriction endonuclease sites located within the DNA sequence encoding mature polypeptides) joined identically to an E. coli trp promoter gave rise to bacterial-produced interferons with distinctly different antiviral activities. The expression plasmid which directs the synthesis of IFN-alpha I was constructed using a gene isolated from a human genomic library in a manner similar to the previous expression of IFN-alpha A and IFN-alpha D cDNA clones. The use of a cell-free transcription-translation system has allowed the calculation of the specific activities of IFN-alpha made from isolated DNA fragments containing these hybrid IFN-alpha genes. These bacterially-derived interferons vary considerably in their ability to inhibit vesicular stomatitis virus (VSV) in different mammalian cells. The results show that the cloned hybrid interferons have unique antiviral activities when compared with the parent interferons, and they demonstrate that more active IFN-alpha s can be made using recombinant DNA techniques.

摘要

利用分子克隆方法构建了4个人源杂交白细胞干扰素（IFN-α）基因，并在大肠杆菌中进行了表达。含有编码人干扰素、IFN-α A、IFN-α D、IFN-α I基因的质粒，以及不同IFN-α基因的几种杂交体（通过在编码成熟多肽的DNA序列内的常见限制性内切酶位点进行体外重组形成），与大肠杆菌色氨酸启动子相同连接后，产生了具有明显不同抗病毒活性的细菌产生的干扰素。指导IFN-α I合成的表达质粒是使用从人基因组文库中分离的基因构建的，其方式类似于之前IFN-α A和IFN-α D cDNA克隆的表达。无细胞转录-翻译系统的使用使得能够计算由含有这些杂交IFN-α基因的分离DNA片段产生的IFN-α的比活性。这些细菌衍生的干扰素在不同哺乳动物细胞中抑制水泡性口炎病毒（VSV）的能力差异很大。结果表明，与亲本干扰素相比，克隆的杂交干扰素具有独特的抗病毒活性，并且表明使用重组DNA技术可以产生更具活性的IFN-α。