CAJ | 학술논문

用农杆菌介导法将嵌合基因GFP-mTn(mTn是微丝结合蛋白Talin的微丝结合域,可以显示活体细胞中微丝的结构)导入蓝猪耳.经激光共聚焦显微镜观察了转基因植株的各种不同组织中融合蛋白的表达和分布情况.在叶片的表皮细胞、保卫细胞、根部的皮层细胞中有融合蛋白的不同程度表达.但仅在保卫细胞中微丝标记状况良好,显示基因表达的组织特异性.经光诱导处于开放态的气孔的保卫细胞微丝呈网状结构,在细胞内无规则分布;经黑暗诱导处于关闭态的气孔保卫细胞中微丝束沿保卫细胞纵轴排列,呈卷曲状分布,并观察到螺旋和环状的微丝结构.在转基因植株的其他部位,例如茎表皮细胞、根毛细胞和花粉粒中,未检测到目的基因的表达.本研究获得的转基因植株为研究气孔运动过程中微丝动态变化提供了有用的材料.
용농간균개도법장감합기인GFP-mTn(mTn시미사결합단백Talin적미사결합역,가이현시활체세포중미사적결구)도입람저이.경격광공취초현미경관찰료전기인식주적각충불동조직중융합단백적표체화분포정황.재협편적표피세포、보위세포、근부적피층세포중유융합단백적불동정도표체.단부재보위세포중미사표기상황량호,현시기인표체적조직특이성.경광유도처우개방태적기공적보위세포미사정망상결구,재세포내무규칙분포;경흑암유도처우관폐태적기공보위세포중미사속연보위세포종축배렬,정권곡상분포,병관찰도라선화배상적미사결구.재전기인식주적기타부위,례여경표피세포、근모세포화화분립중,미검측도목적기인적표체.본연구획득적전기인식주위연구기공운동과정중미사동태변화제공료유용적재료.
By transformation mediated by Agrobacterium tumefacien, we successfully transferred the chimeric gene of GFP-mTn ( mTn is the binding domain of microfilament binding protein talin from mouse, which can show the microfilament in living cell ) into Torenia fournieri. Using confocal laser scanning microscopy (CLSM), the distribution of fusion protein in different kinds of tissues and cell in transgenic Torenia fournieri was observed. GFP fluorescence was found in leaf epidermal cell, stomatal guard cell and root epidermal cell. Actin filaments can be visualized clearly only in guard cells. In the guard cells of open stomata under light, actin filaments arrange reticularly and randomly in cortical cytoplasm. In the guard cells of closed stomata under darkness, actin filaments arrange curly along the longitude of guard cell, and some helix and ring structures were found. GFP fluorescence was not found in other cell types, including stem epidermal cell, root hair cell and reproductive organs. The transgenic Torenia fournieri we got provides a suitable material to study dynamics of actin filament in stomatal guard cell.