黄瓜花叶病毒2b基因瞬时表达载体的构建及鉴定
Subcloning of CMV 2b gene and the construction of the transient expression vector
黄瓜花叶病毒（CMV）2b蛋白由CMV的亚基因组RNA4A表达产生，是最早被证实为抑制转录后基因沉默的抑制子之一。从三种不同CMV的pFRNA2中克隆出2b基因片段，将其PCR产物与pUC18载体连接，经PCR鉴定，测序比对正确后，用SacI/BamHI双酶切得到2b基因片段，亚克隆至经相同双酶切的pBI121载体。经2b基因片段的PCR鉴定，筛选出阳性重组体，即为三种pBI121-2b瞬时表达载体。将载体电转化到农杆菌EHA105中，通过与携带pBI121-GFP的菌液共浸润分析各2b蛋白的生物学活性。为研究2b基因的致病性与抑制转录后基因沉默的内在关系提供必要的生物学材料。
MV 2b protein were translated from the subgenomic RNA(RNA4A) of RNA2, and was one of the first determined RNA silencing suppressors. The different 2b gene fragments were cloned in pUC18 from different pF209 of Rad35-CMV, Rad352b 164-CMV, Fny-CMV, then subcloned in PBI121 transient expression vector. After verification by restriction analysis, PBI121-2b transient expression vector containing about 300bp 2b gene fragment was successfully constructed by means of recombinant DNA technology. These results could be used for further research of 2b gene function and mechanism of RNA silencing.
李喜英、赵志敬、杜志游、陈集双
分子生物学遗传学生物工程学
黄瓜花叶病毒2b蛋白瞬时表达载体基因沉默
MVMV2bransient expression vectorGene silencing
李喜英,赵志敬,杜志游,陈集双.黄瓜花叶病毒2b基因瞬时表达载体的构建及鉴定[EB/OL].(2010-01-28)[2025-08-10].http://www.paper.edu.cn/releasepaper/content/201001-1255.点此复制
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