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采用GUS标记技术研究钙调素在转基因烟草中的分布

Calmodulin Distribution in the Transgenic Tobacco Using GUS Reporter Gene

  • 摘要: 为了监测钙调素(CaM)在植物细胞内的分布并探索其生物学功能,将水稻CaM和GUS融合基因(cam·gus)分别置于35S启动子和花粉特异启动子LAT52-7控制下,构建出载体pMD/CaM·GUS和pBI/LAT·CaM·GUS,转化烟草(Nicotianatobacumcv.SRⅠ),GUS染色结果显示,CaM·GUS融合蛋白广泛分布于植物根、茎、叶等组织,根尖生长点细胞、根毛细胞、表皮毛细胞、气孔保卫细胞、维管束细胞的细胞质中融合蛋白含量较多.花粉粒中也分布着CaM,并在萌发沟处表现出较高的浓度.保卫细胞和表皮毛细胞中CaM分布在相邻细胞连接处细胞模内侧,呈现出极性分布.研究结果表明,细胞内CaM的不均匀分布可能与细胞功能相关.

     

    Abstract: In order to study the localization of calmodulin in the plant cell and it's biological function,gene constucts containing the fusiongene(cam·gus)under the control of the 35S CaMV promoter or the pollen specific promoter LAT 52-7 from tomato,were in troduced into Nicotianatobacum cv.SRI by leaf disctrans for mation respectively.GUS reaction show that in the tran sgenicline pMD CaM·GUS,the fusion protein CaM·GUS was widely observed in roots,stems and leaves.And it had a higher levels throughout the cytosal of root meristem region,quiescence center cell,root hair cells,epidermal cells,and vascular cells.CaM was also detected in polar distributionsty leunder the membrane of the jointsite between-guard cells.Inp BILAT·CaM·GUStran sgenic line,CaM only localized in the cytop last of pollen grains,and a higher expression of CaM was detected at germination zones.Our results show that Ca Mex hibitsa symmetric distribution the plant cells,which has a very tight relation ship with cell biological function.

     

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