蜻蜓凤梨AfAG基因的克隆、载体构建及遗传转化

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本研究以蜻蜓凤梨为材料,利用RACE技术获得一个与拟南芥AGAMOUS(AG)同源的编码基因c DNA,并将其命名为Af AG。Af AG基因的c DNA全长1 422 bp,开放阅读框为723 bp,可翻译成240个氨基酸。利用NCBI对Af AG蛋白进行序列分析,结果显示Af AG蛋白含有MADS-box蛋白家族共有的功能结构域:MADS和K-BOX两个结构域。与水稻(Os AG);玉米(Zm AGL);拟南芥(At AG);菜心(Br AGL);椰子(Cn AGL);白兰花(Ma AGL)的同源性比对结果分别为65%、63%、68%、65%、83%、75%。本研究成功构建了Ca MV35S-Af AG过表达载体,用农杆菌介导的方法将表达载体成功转入到拟南芥中,为进一步研究蜻蜓凤梨开花分子机理提供了理论依据。 In this study, dragonfly pineapple as material, using RACE technology to obtain a homologue of Arabidopsis AGAMOUS (AG) encoding gene cDNA, and named Af AG. The cDNA of Af AG gene is 1 422 bp in length and 723 bp in open reading frame, which can be translated into 240 amino acids. Sequence analysis of Af AG protein using NCBI showed that Af AG protein contained two functional domains shared by MADS-box protein family: MADS and K-BOX. The results of homology comparison with Os AG, Zm AGL, At AG, Br AGL, Cn AGL and Ma AGL were 65 %, 63%, 68%, 65%, 83%, 75%. In this study, Ca MV35S-Af AG overexpression vector was successfully constructed. The expression vector was successfully transferred into Arabidopsis by Agrobacterium-mediated transformation, which provided a theoretical basis for further study on the flowering molecular mechanism of dragonfly pineapple.
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