腊梅花水通道蛋白CpTIP基因甘露糖筛选体系植物表达载体的构建

[目的】构建以甘露糖为筛选剂的抗旱、抗盐碱植物表达载体，进一步选育无标记的抗逆作物品种。[方法]利用腊梅花水通道蛋白CpTIP cDNA和大肠杆菌pmi基因构建植物表达载体，将抗逆基凶与甘露糖正向选择系统结合。pmi基因植物表达载体（pPMI）的构建：用XhoI酶切植物表达载体pCAMBIA2301，切除Kan基因，并用CIP酶进行去磷酸化，然后与XhoI酶切后PCR产物连接转化。腊梅花水通道蛋白CpTIP基因甘露糖筛选体系植物表达载体（pPMI：：CpTIP）的构建：用和nI和Xbal酶切pPMI植物表达载体和克隆载体，并将酶切后pmi植物表达载体与腊梅花水通道蛋白CpTIP基因连接转化。转化鉴定均按Sambrook等的方法进行。[结果]成功构建了腊梅花水通道蛋白CpTIP基因甘露糖筛选体系植物表达载体pPMI：：CpTIP。[结论]所构建的载体结合了抗逆基凶和忖露糖正向选择系统的优点，将抗逆基因与环境友好型筛选体系很好的结合起来。

Construction of Chimonanthus praecox(L.)Link Aquaporin CpTIP Plant Expression Vector with Mannose Selection System

[ Objective] The aim was to construct drought and saline-alkaline resistance plant expression vector with mannose as selective agent, and further breed unmarked resilient varieties. [ Method] The plant expression vector was constructed by using Chimonanthus praecox（ L. ）Link aquapor.in CpTIP cDNA and Escherichia coli pmi gene, combined stress resistance gene with mannose positive selection system. [ Result] The test successfully constructed the plant expression vector pPMI：：CpTIP. [ Conclusion] The constructed vector linked advantages of stress resistance gene and mannose positive selection system.