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Position: Home > Articles > RT-PCR of 2b gene fragments of some CMV isolates from China and comparison with their sequences Acta Phytopathologica Sinica 2003,33 (2) 146-150

我国不同CMV分离物2b基因片段的RT-PCR扩增及其序列比较

作  者:
庄木;王晓武;谢丙炎;郑文光;冯兰香;方智远
单  位:
中国农业科学院蔬菜花卉研究所生物技术室
关键词:
黄瓜花叶病毒;2b基因;反转录-聚合酶链式反应;转录后基因沉默
摘  要:
本研究根据黄瓜花叶病毒2b基因的保守序列设计引物,利用一步RT-PCR技术对多个不同寄主和不同地理来源的黄瓜花叶病毒分离物的2b基因片段进行扩增,获得了含该基因全长约90%的cDNA片段(300bp)。序列测定与比较分析结果表明,国内不同CMV分离物2b基因片段核酸序列同源性达93%以上,推测的氨基酸序列同源性超过90%,且均属于亚组Ⅰ。
译  名:
RT-PCR of 2b gene fragments of some CMV isolates from China and comparison with their sequences
作  者:
ZHUANG Mu;WANG Xiao-wu;XIE Bing-yan;ZHENG Wen-guang;FENG Lan-xiang, FANG Zhi-yuan Biotechnology Department, Institute of Vegetables and Flowers, CAAS, Beijing 100081, China
关键词:
CMV;;2b gene;;RT-PCR;;PTGS
摘  要:
A simple and rapid method for detecting Cucumber mosaic virus (CMV) was developed. CMV 2b gene fragments (300 bp) of 9 different isolates were successfully amoplified by one step RT-PCR from virus samples prepared by grinding in an extraction buffer. RT-PCR products of 9 isolates were cloned into pMD18-T vector and 7 of them were sequenced. The comparison of these sequences revealed that 2b gene fragments of different CMV isolates were highly conservative, and the identity of nucleotide and amino acid sequences were over 93% and 90%, respectively. It showed that all the Chinese iso- lates used belong to the subgroup Ⅰ of CMV.

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