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马铃薯卷叶病毒(PLRV)内蒙古分离物基因间隔区(IS)的克隆与序列分析

DOI: 10.7668/hbnxb.2007.06.005, PP. 19-23

Keywords: 马铃薯卷叶病毒(PLRV),内蒙古分离物,基因间隔区(IS),cDNA克隆,序列分析

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Abstract:

根据Genbank已报道马铃薯卷叶病毒(PLRV)基因组序列,分析其基因间隔区(IS)两端的保守区,自行设计、合成一对特异性引物,以PLRV内蒙古分离物总RNA为模板,经RT-PCR扩增得到含PLRVIS的一段369bp的cDNA,克隆于载体pBS-T中。重组质粒经PCR鉴定、酶切分析和核苷酸序列测定,并进一步与PLRV其他分离物的同源序列作比对。结果表明:克隆的PLRV内蒙古分离物的IS序列与其他全部已发表的13个全基因组中的IS核苷酸序列有很高的同源性,最高达到100%,平均为97.90%,高于这13个PLRV全基因组序列96.81%的同源性,说明IS序列不仅在PLRV的不同株系间比较保守,而且在PLRV的全基因组序列中也是相对保守的。研究结果预示,将IS构建成RNA干扰型结构导入马铃薯,将有可能获得抗PLRV多种株系且抗性更高的转基因植株。

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