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口蹄疫病毒具有生物活性的蛋白酶是通过在大肠杆菌中克隆的病毒cDNA表达的。

Biologically active protease of foot and mouth disease virus is expressed from cloned viral cDNA in Escherichia coli.

作者信息

Klump W, Marquardt O, Hofschneider P H

出版信息

Proc Natl Acad Sci U S A. 1984 Jun;81(11):3351-5. doi: 10.1073/pnas.81.11.3351.

Abstract

Foot and mouth disease virus O1K cDNA had been cloned in Escherichia coli. Here we report on in vitro recombination of cDNA fragments according to the cDNA restriction map and on expression of viral proteins in E. coli. Use was made of the expression vector pPLVP1 , which is known to express the virus capsid protein VP1. Recombined cDNAs of various sizes were inserted downstream from the VP1 gene. The constructed plasmids differ from each other in the number of virus genes coding for nonstructural proteins. The effects caused by their expression in E. coli are compared. It is shown that the virus protease is expressed in E. coli as an active enzyme that recognizes the simultaneously produced virus-specific polyprotein as substrate. The virus protease gene was mapped 5'-adjacent to the virus replicase gene.

摘要

口蹄疫病毒O1K的互补脱氧核糖核酸(cDNA)已在大肠杆菌中克隆出来。在此,我们报告根据cDNA限制性图谱进行的cDNA片段体外重组,以及病毒蛋白在大肠杆菌中的表达情况。使用了表达载体pPLVP1,已知该载体可表达病毒衣壳蛋白VP1。将各种大小的重组cDNA插入VP1基因下游。构建的质粒在编码非结构蛋白的病毒基因数量上彼此不同。比较了它们在大肠杆菌中表达所产生的影响。结果表明,病毒蛋白酶在大肠杆菌中作为一种活性酶表达,该酶将同时产生的病毒特异性多蛋白识别为底物。病毒蛋白酶基因定位于病毒复制酶基因5'相邻处。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/61a7/345505/7509b1f33cec/pnas00612-0099-a.jpg

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