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- W2038611282 abstract "Late embriogenesis abundant (LEA) protein is one of the hot topics in plant stress physiology. In this study, an RNAi expression vector harboring MsLEA3-1 gene fragment from alfalfa (Medicago sativa L.) was constructed. Two pairs of specific primers with different enzyme sites were designed based on the sequence of MsLEA3-1 (GenBank accession number EU665182). With the template of PMD-LEA plasmid constructed, positive-sense strand and antisense strand were obtained, which were separately inserted into the expression vector pART27. A hairpin structure in the RNAi vector pART-F-R was confirmed by the digestion of restriction enzymes. pART-F-R was transformed into tobacco via Agrobacterium-mediated transformation system, and 16 transgenic plants were obtained after PCR validation. 根据紫花苜蓿LEA蛋白MsLEA3-1基因(GenBank登录号为EU665182)序列, 设计两对含有酶切位点的特异性引物LEAf1/ LEAf2和LEAr1/LEAr2, 以构建好的PMD-LEA质粒为模板, 分别合成用于构建干扰载体的正反义片段pMD-F和pMD-R, 将正反义片段分别插入表达载体pART27的相应位置, 构建成含有发夹结构的RNAi载体pART-F-R, 经过Not I酶切鉴定, 证明载体构建成功。通过农杆菌介导的方法, 以干扰表达载体pART-F-R转化烟草, 经过PCR检测, 得到16株阳性转基因植株, 为MsLEA3-1基因的功能研究奠定基础。" @default.
- W2038611282 created "2016-06-24" @default.
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- W2038611282 date "2010-09-01" @default.
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- W2038611282 title "Construction of ihpRNA Expression Vector of MsLEA3-1 from Medicago sativa L. and Genetic Transformation in Tobacco" @default.
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- W2038611282 doi "https://doi.org/10.1016/s1875-2780(09)60073-0" @default.
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