Construction and characterization of two infectious molecular clones of encephalomyocarditis virus.
Construction and characterization of two infectious molecular clones of encephalomyocarditis virus.
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脑心肌炎病毒两个感染性分子克隆的构建和表征。
DOI：
10.1128/jvi.64.2.913-917.1990
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发表时间：
1990
影响因子：
5.4
通讯作者：
Jordan,GW
中科院分区：
文献类型：
--
作者：
Naviaux,RK;Cohen,SH;VandenBrink,KM;Jordan,GW
我们构建了两个脑心肌炎(EMC)病毒感染性分子克隆并对其进行了鉴定。这两个构建物，PDL和PDA，都是由来自EMC病毒的糖尿病变异体(EMC-D)的五个重叠的cDNA克隆和两个合成的寡核苷酸盒组装而成的。PDA在衣壳蛋白1AB的1720位含有单点突变，该衣壳蛋白来源于EMC病毒(EMC-B)的非糖尿病变异体。该点突变导致精氨酸(EMC-B)的氨基酸取代赖氨酸(EMC-D)。我们的构建说明了两个新的发现：(I)稳定克隆EMC病毒长Poly(C)区域的问题可以通过使用缩短的、合成的聚(DC-DG)寡核苷酸盒来规避，以及(Ii)衣壳蛋白1AB泡发区的单点突变导致其电泳迁移率的变化和重组病毒的空斑大小的变化。
We constructed and characterized two infectious molecular clones of encephalomyocarditis (EMC) virus. Both constructs, pDL and pDA, were assembled from five overlapping cDNA clones derived from the diabetogenic variant of EMC virus (EMC-D) and from two synthetic oligonucleotide cartridges. pDA contained a single point mutation at position 1720 within the "puff" region of capsid protein 1AB that was derived from the nondiabetogenic variant of EMC virus (EMC-B). This point mutation resulted in an amino acid substitution of arginine (EMC-B) for lysine (EMC-D). Our construction illustrates two novel findings: (i) that the problem of stably cloning long poly(C) tracts of EMC virus can be circumvented by the use of a shortened, synthetic, poly(dC-dG) oligonucleotide cartridge, and (ii) that a single point mutation in the puff region of the capsid protein 1AB leads to change in its electrophoretic mobility and to a change in the plaque size of recombinant virus.