双元植物表达载体pCAMBIA2300-HaBADH-HaCMO的构建及转化  

Construction and transformation of the plant binary expression vector pCAMBIA2300-HaBADH-HaCMO

作　　者：石磊 周晓燕[1,2] 甘晓燕 马洪爱 宋玉霞 

机构地区：[1]宁夏农业生物重点实验室,宁夏银川750002 [2]宁夏大学生命科学学院,宁夏银川750021

出　　处：《西北农林科技大学学报（自然科学版）》2012年第6期210-216,223,共8页Journal of Northwest A&F University(Natural Science Edition)

基　　金：宁夏回族自治区科技攻关项目(KGZ-16-07-02)

摘　　要：【目的】构建双元植物表达载体,通过遗传转化提高植物的抗逆能力。【方法】将从超旱生、耐盐植物梭梭(Haloxylon ammodendron)中克隆得到的HaBADH基因,定向导入植物表达载体pCAMBIA2300-35S-OCS中,以HaCMO替换pCAMBIA2300-HaBADH中的抗性基因NPTⅡ,构建双元植物表达载体pCAMBIA2300-HaBADH-HaCMO,并通过农杆菌介导法将其转入粳稻品种"优引三号",对所获得的转基因植株进行PCR检验。【结果】成功构建了双元植物表达载体pCAMBIA2300-HaBADH-HaCMO,并获得了携带有pCAMBIA2300-HaBADH-HaCMO的水稻阳性植株5株,经PCR检测,转入成功。【结论】将梭梭抗逆基因HaBADH和HaCMO构建到一个表达载体中,并成功转化水稻,为转入基因后水稻植株内甜菜碱合成和积累过程的深入分析提供了条件。【Objective】 The study was done to construct a binary expression vector to improve stress-tolerance of target plants by genetic transformation.【Method】 We introduced HaBADH directly into plant expression vector pCAMBIA2300-35S-OCS,and replaced NPTⅡ with HaCMO,and constructed a binary expression vector.HaBADH and HaCMO were both cloned by our laboratory and pCAMBIA2300-HaBADH-HaCMO was transformed into rice Youyin-3 by Agrobacterium-mediated transformation and transgenatic plants were detected with PCR.【Result】 We constructed the plant binary expression vector pCAMBIA2300-HaBADH-HaCMO successfully,and obtained 5 transgenic rice plants which contained the vector by PCR analysis.【Conclusion】 This study introduces HaBADH and HaCMO to pCAMBIA2300-35S-OCS,and constructs a binary expression vector pCAMBIA2300-HaBADH-HaCMO,and transforms it to rice,which prepares for study synthesis and accumulation of glycine betaine in transgenetic rice.

关 键 词：梭梭 甜菜碱醛脱氢酶 胆碱单加氧酶 

分 类 号：Q782[生物学—分子生物学]