Foreign complementary sequences facilitate genetic RNA recombination in brome mosaic virus.
Foreign complementary sequences facilitate genetic RNA recombination in brome mosaic virus.
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外来互补序列促进雀麦花叶病毒中的遗传 RNA 重组。
DOI：
10.1006/viro.1995.1163
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发表时间：
1995
期刊：
影响因子：
--
通讯作者：
Bujarski,JJ
中科院分区：
文献类型：
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作者：
Dzianott,A;Flasinski,S;Bujarski,JJ
我们已经证明了局部反义序列可以介导溴花叶病毒（BMV） rna的3 '非编码区基因重组（P. Nagy and J. J. Bujarski, 1993, Proc, Natl）。学会科学。美国90,6390-6394)。在这里，我们发现外来互补插入可以指导内部区域内BMV RNA3组分之间的交叉。从豇豆叶绿体斑驳病毒（CCMV） RNA3中提取的170 nt多核苷酸以正义或反义方向插入BMV外壳蛋白开放阅读框起始带的上游。当分别接种在局部病变寄主的叶片上接种BMV时，得到的突变体BCC+和BCC-保持不变的CCMV插入。相比之下，当接种含有两种突变RNAs3的混合物时，相当一部分病变积累了缺乏CCMV插入物的BMV RNA3。3 '标记突变的存在证实了BMV RNA3后代是由于互补序列中BCC+和BCC-之间的交叉而产生的。当RNA混合物在宿主植物上接种前退火时，观察到重组出现的频率最高。我们的结果证实了一个概念，预测了RNA病毒中异源双工介导的重组功能的一般性质。讨论了该方法在重组RNA技术中的应用实例。
We have demonstrated that local antisense sequences can mediate genetic recombination within the 3′ noncoding region among brome mosaic virus (BMV) RNAs (P. Nagy and J. J. Bujarski, 1993, Proc, Natl. Acad. Sci. USA 90, 6390-6394). Here we show that foreign complementary inserts can direct crossovers between BMV RNA3 components within an internal region. A 170-nt polynucleotide derived from the cowpea chlorotic mottle virus (CCMV) RNA3 was inserted just upstream of the initiation cordon of the BMV coat protein open reading frame in either sense or antisense orientations. The resulting respective mutants, BCC+ and BCC-, maintained unchanged CCMV inserts when inoculated separately on leaves of a local lesion host for BMV. In contrast, when a mixture containing both mutated RNAs3 was inoculated, a significant fraction of lesions accumulated the BMV RNA3 lacking the CCMV insert. The presence of a 3′ marker mutation confirmed that the BMV RNA3 progeny arose due to crossovers between BCC+ and BCC- within the complementary sequences. The highest frequency of recombinant appearance was observed when the RNA mixtures were annealed prior to inoculation on the host plants. Our results confirm a concept predicting the general nature of the heteroduplex-mediated recombination functioning in RNA viruses. Examples of possible applications of this approach in recombinant RNA technology are discussed.