摘要
高效易用的植物表达载体在基因功能相关研究中起着重要的作用。以植物表达载体p BA002为基础载体,使用抗草甘膦CP4-EPSPS基因替换原载体的Bar筛选标记基因,获得新的植物表达载体p ES002。将大豆耐逆相关Trihelix类转录因子Gm GT-2A连接到p ES002,构建了同时含耐逆相关基因和抗除草剂标记基因的植物表达载体p ES002-Gm GT-2A。利用农杆菌介导的花序浸润法转化拟南芥,并对转基因植株进行抗草甘膦和耐盐性鉴定。结果表明,转基因植株在抗草甘膦和耐盐性上均显著强于对照,表明该载体可以用于转基因相关的基因功能研究。
An efficient and easy-to-use plant expression vector plays an important role in the research of gene func-tions.In this study, the glyphosate resistant gene CP4-EPSPS was cloned to replace the original selective marker Bar gene of plant expression vector pBA002.The new plant expression vector was named as pES002.A Trihelix tran-scription factor GmGT-2A was subsequently ligated into pES002 to form a new plant expression vector, namely pES002-GmGT-2A.This vector was transformed into Arabidopsis by floral dipping method to verify the functions of both CP4-EPSPS and GmGT-2A genes.Results showed that the transgenic lines greatly improved the tolerance to both glyphosate and high salinity stress, indicating that both the new selective marker gene and the target gene worked well.Thus, this vector was successfully constructed and could be further used in transgenic related research.
出处
《浙江农业学报》
CSCD
北大核心
2015年第2期133-140,共8页
Acta Agriculturae Zhejiangensis
基金
转基因生物新品种培育重大专项(2012ZX08009004,2013ZX08004002)
关键词
草甘膦
转录因子
表达载体
CP4-EPSPS
GmGT-2A
CP 4-EPSPS
GmGT-2A
glyphosate
transcription factor
expression vector
