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玉米叶绿体表达载体构建及绿色荧光蛋白基因瞬时表达检测

DOI: 10.7668/hbnxb.2008.04.017, PP. 72-76

Keywords: 玉米,叶绿体表达载体,瞬时表达,GFP荧光

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Abstract:

Tps1是生物保护物质海藻糖的关键合酶基因。构建含Tps1的玉米叶绿体表达载体,首先从玉米叶绿体基因组中克隆16S/trnI-trnA/23S片段,作为定点整合同源片段;然后将编码酵母海藻糖合酶基因Tps1表达盒(Prrn-Tps1-TpsbA-ter)、草丁膦抗性基因Bar表达盒(RpsbApro-Bar-RpsbAter)、卡那霉素抗性基因NptII和绿色荧光蛋白基因Gfp构建的融和基因表达盒(Prrn-NptII/Gfp-RrbcLter)一起克隆到定点整合同源片段之间,构建了玉米叶绿体的稳定表达载体mTKGB。通过基因枪轰击法将mTKGB转化到玉米幼嫩叶片中,培养2d后,在激光扫描共聚焦显微镜下观测到玉米一些细胞的叶绿体中有很强的GFP绿色荧光,结果表明构建的载体可在玉米叶绿体中表达。

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