拟南芥MAP65-3和SEC6的互作及其在胞质分离中的作用
毕业论文关键词：MAP65-3；SEC6；胞质分离
Map65-3 Interacts with Sec6 and Their Function during Cytokinesis in Arabidopsis
Abstract：Cytokinesis is a significant biological process in cell pision in the ahead of the establishment of cell plate. Previous researches indicated that the establishment of cell plate is involved in vesicle tethering and assistance of microtubule bundles to locate. Tethering factor-Exocyst assists vesicle to recognize and fuse with targeted plasma membrane. SEC6, a subunit of Exocyst, is located in cell plate, which have a great impact on cytokinesis in plants. The stability of microtubule bundles’ shape and structure is maintained by MAPs, whose family member-MAP65-3 used to lie where cell pision is very frequent and it takes part in establishment of cell wall. Our research showed that MAP65-3 interacts with SCE6，mutation of map65-3 or sec6 in Arabidopsis will cause stunted plants and defective seeds, which also has an effect on the distribution of protein-KNOLLE during the cytokinesis.
Key words: MAP65-3；SEC6；cytokinesis
目 录
摘要4
关键词4
Abstract4
Key words4
引言4
1材料与方法6
1.1植物材料 6
1.2菌株和质粒 6
1.3实验试剂7
1.3.1常用试剂配方7
1.3.2培养基的配制7
1.4 植物总DNA的提取8
1.5 植物总RNA的提取8
1.6 反转录PCR8
1.7 重组载体的构建9
1.8 引物设计及PCR扩增9
1.9转化实验9
1.9.1转化大肠杆菌9
1.9.2转化农杆菌9
1.10质粒共转酵母双杂交系统9
1.10.1 重组载体构建9
1.10.2 酵母感受态制备9
1.10.3 重组载体共转酵母10
1.11 双分子荧光互补10
1.11.1 重组载体构建10
1.11.2 农杆菌侵染液配制10
1.12 拟南芥幼苗根尖的FM4-64染色10
2 结果与分析10
2.1 MAP65-3和SEC6存在互作关系10
2.1.1 酵母双杂交10
2.1.2 双分子荧光互补11
2.2 SEC6在map65-3 中的定位没有受到影响12
2.3 拟南芥map65-3和PRsec6突变体对生理功能的影响12
3讨论13
3.1MAP65-3在拟南芥胞质分离过程中发挥重要作用13
3.2 MAP65-3和SEC6在拟南芥胞质分离过程中的互作关系及其生理学意义13
致谢13
参考文献14
拟南芥MAP65-3和SEC6的互作及其在胞质分离中的作用
引言
自然界中，细胞分裂是各种生物体生长、发育、繁殖的主要生物学途径。细胞质分裂是细胞有丝分裂的最后一步，对于细胞增殖的保真性不可或缺。该过程伴随细胞板的形成，受到囊泡定向运输的协助。细胞板的基本形成过程大致如下，首先由高尔基体衍生而来的囊泡迎着成膜体移动到达细胞的等分区-赤道板上，而后囊泡之间相互融合成管状囊泡网，管状囊泡网拓展连接、膨胀形成富含胼胝质的光滑薄孔网状结构管状网络，细胞板以离散的方式向母细胞壁延伸，并在延伸边缘形成许多指状融合管；当指状融合管延伸到达母细胞皮层时，与之发生融合；最后细胞板上网孔被纤文素等物质填充，细胞板成熟产生新生细胞壁[1]。从细胞板的形成过程可知：囊泡的定向运输对于细胞板的正确形成起到了至关重要的作用。