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[脊髓灰质炎病毒3C蛋白酶基因在大肠杆菌细胞中的克隆与表达]

[Cloning and expression of the 3C-proteinase gene from the poliomyelitis virus in E. coli cells].

作者信息

Sablina E P, Amerik A Iu, Antonov V K

出版信息

Bioorg Khim. 1991 Sep;17(9):1201-12.

PMID:1666949
Abstract

Expression of the 3C protease gene of poliovirus type 1 (Mahoney) in E. coli cells using various vectors was studied. The 3C gene was shown to be expressed effectively upon its cloning in HindII/HindII (bases 5240 to 6770) and in HindII/HindIII (bases 5240 to 6056) fragments of poliovirus cDNA in pTTQ8 vector containing tac-promoter and lacI-repressor gene. Products of processing at the N-terminal 3C protease Gln-Gly site and polypeptides formed upon translation from an alternative methionine, which was coded by bases 5516-5518 of poliovirus cDNA, were found among virus-specific proteins. Processing at the C-terminal 3C protease Gln-Gly site was not observed.

摘要

研究了使用各种载体在大肠杆菌细胞中表达1型脊髓灰质炎病毒(Mahoney株)的3C蛋白酶基因。当3C基因克隆到含有tac启动子和lacI阻遏基因的pTTQ8载体中的脊髓灰质炎病毒cDNA的HindII/HindII(碱基5240至6770)和HindII/HindIII(碱基5240至6056)片段中时,显示其能有效表达。在病毒特异性蛋白中发现了在N端3C蛋白酶Gln-Gly位点的加工产物以及由脊髓灰质炎病毒cDNA的碱基5516 - 5518编码的另一个甲硫氨酸翻译形成的多肽。未观察到在C端3C蛋白酶Gln-Gly位点的加工。

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