DEVELOPMENT OF A RECOMBINANT RNA TECHNIQUE FOR THE CONSTRUCTION OF CHIMERIC RNA WITH A LONG POLY(C) TRACT

被引:11
作者
BAE, YS
KANG, Y
OHTSUKA, E
YOON, JW
机构
[1] UNIV CALGARY,FAC MED,JULIA MCFARLANE DIABET RES CTR,3330 HOSP DR NW,CALGARY T2N 4N1,AB,CANADA
[2] UNIV CAGARY,FAC MED,DEPT MICROBIOL & INFECT DIS,CALGARY T2N 4N1,AB,CANADA
[3] HOKKAIDO UNIV,FAC PHARMACEUT SCI,SAPPORO,HOKKAIDO 060,JAPAN
基金
英国医学研究理事会;
关键词
D O I
10.1093/nar/21.11.2703
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The murine cardioviruses and bovine aphthoviruses are distinguished from other (+) strand RNA viruses by their long poly(C) tract in the 5'-noncoding region. The presence of this poly(C) tract has long hampered the construction of full-length cDNA with the complete poly(C) tract, because long poly(dC-dG) homopolymer-containing plasmids are difficult to amplify in bacterial systems. To overcome this problem, we constructed a chimeric RNA by joining the poly(C) region of the viral RNA to the 5'-truncated RNA transcript of the encephalomyocarditis (EMC) virus cDNA. The non-chimeric, recombinant EMC virus with a short poly(C) tract produces recombinant progeny virus, but this is not pathogenic in vivo. On the other hand, the EMC viral RNA chimera with the complete poly(C) tract produces recombinant progeny virus that is pathogenic in vivo. This method of viral RNA construction will be invaluable for functional studies of other cardioviruses and aphthoviruses, as well as for recombinant RNA manipulations.
引用
收藏
页码:2703 / 2708
页数:6
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