磷脂酰肌醇合成酶PIS基因植物表达载体的构建

Construction of plant expression vector of phosphatidyliositol synthase gene PIS

磷脂酰肌醇(PI)是真核细胞中主要的磷脂组分,磷脂酰肌醇合成酶(PIS)是磷脂酰肌醇合成途径中的关键酶,它的功能是催化myo-肌醇和CDP-DG反应产生磷脂酰肌醇。将玉米PIS基因与载体pCAMBIA2300-35s连接,得到植物表达载体pCAMBIA2300-35s-PIS,该载体带有CaMV35s启动子、PIS基因、终止序列及一个CaMV35s启动子启动的NPTⅡ抗性筛选基因。将构建好的PIS植物表达载体转入农杆菌中,用于将来国槐等木本植物的转化。

Phosphoinosidtide（PI） is a major phospholipid in eukaryotie cells. Phospholipid signaling pathway （PI pathway） plays a key role in plant growth, development and responses to environmental stress conditions. Phosphatidyliositol synthase （PIS） is a key enzyme in the phosphatidyliositol synthesis pathway, and can synthesize phosphoinosidtide with CDP--DG and myo--inositol. The gene PIS was cloned into the reconstructed vector pCAMBIA2300--35s. The vector pCAMBIA2300--35s --PIS contained CaMV35s promoter, PIS gene, terminator sequence and NPT Ⅱ selectable marker. The sequence of the vector was conformed by enzyme cutting and order--checking. The vector pCAMBIA2300-PIS was transformed into the Agrobactri- um tumefaeiens GV3101 and LBA4404 used in the genetic transformation of the xylophyta.