生长素结合蛋白与纤维素合成酶在同一细胞中的标记与定位  

Labeling and location of ABP1 and CesA1 in the same tobacco plant cell

作　　者：陈宗星[1] 刘晓柱[1] 黄妤[1] 黄丽华[1] 张学文[1] 

机构地区：[1]湖南农业大学生物科学技术学院,湖南长沙410128

出　　处：《湖南农业大学学报（自然科学版）》2012年第6期602-606,F0003,共6页Journal of Hunan Agricultural University(Natural Sciences)

基　　金：北京大学国家蛋白质与基因工程重点实验室开放项目

摘　　要：生长素通过其结合蛋白(ABP1)促进细胞纤维素合成并使细胞伸长生长。为探明ABP1与纤维素合成酶(CesA1)这2种蛋白质之间的相互关系,分别对这2种蛋白质进行了荧光标记,构建了绿色荧光蛋白(GFP)对ABP1融合标记以及青色荧光蛋白(CFP)对CesA1融合标记的2种载体。采用烟草叶片侵注法,将2种重组体转入烟草叶片同一细胞进行瞬时表达。扫描激光共聚焦显微成像观察到转基因瞬时表达的烟草铺列细胞的细胞膜上有强烈两色荧光信号,表明这2种蛋白集中在细胞膜上分布,其分布区域存在明显的重叠。在细胞的内膜系统中绿色荧光信号相对较强,青色荧光信号较弱,表明内膜系统中也分布着一定量的ABP1,而纤维素合成酶在内膜系统中不会集中分布。在叶片铺列细胞间嵌合交错区可以观察到青色荧光的高亮点,表明此处存在纤维素合成酶的高密度分布,也说明在此部位有更多的纤维素合成。The auxin binding protein （ABP1） is involved in the response to auxin signal to induce the elongation of plant cell. In order to understand the interaction between ABP1 and CesA1, two chimera expression vectors are constructed to label ABP1 with GFP and CesA1 with CFP respectively. Two reeombinants are transformed into tobacco leaf for transient expression in the same epidermal cell via infiltration. Green and cyan florescent signals are observed in the same pavement cells of the infiltrated leaf region by confocal microscope analysis, indicating proteins ABP1 and CesA1 are accumulated and overlaying distributed in the cell membranes. But in the endomembranes, the green signals are strong while the cyan signals are week, demonstrating that it is ABP 1 but not CesA 1 which is also collectively distributed in the endomembranes system. Cyan florescent signals are observed in interdigitation area of the pavement cells, which indicates intensive distribution of CesAl and more celluloses were synthesized in this area.

关 键 词：生长素结合蛋白 纤维素合成酶 荧光标记 细胞定位 

分 类 号：Q71[生物学—分子生物学]