拟南芥PTR3基因过表达载体构建及转基因植株的获得
Construction of PTR3 Over-expression Carrier and Identification of Transgenic Lines in Arabidopsis thaliana
管灵霞 1樊婷婷 1倪娇娇 1马文佳 1曹树青1
作者信息
- 1. 合肥工业大学生物与食品工程学院,安徽合肥230009
- 折叠
摘要
[目的]构建PTR3基因的过表达栽体,并建立PTR3基因过表达植株,为进一步研究PTR3基因在调控重金属胁迫应答中的功能提供理论依据.[方法]提取野生型拟南芥总mRNA,并以反转录的cDNA为模板扩增出PTR3基因CDS全长,通过限制性核酸内切酶酶切、T4DNA连接酶连接,将该基因连接到带有35S启动子的pBI121载体上.[结果]将重组载体转化至农杆菌GV3101菌株,通过浸花法将重组载体转化到野生型植株中,通过抗性筛选和遗传鉴定获得纯合的PTR3转基因阳性植株.[结论]PTR3过表达载体构建成功并获得转基因阳性植株,为研究PTR3基因的分子功能奠定基础.
Abstract
[Objective] The aim was to construct PTR3 over-expression vector and generate PTR3 over-expression transgenic lines,provide basis for further research functions of PTR3 in response to heavy netal stress.[Method] Total mRNA was extracted from wild Arabidopsis thaliana and employed in a reverse transcription.The CDS fragment of PTR3 gene was amplified by PCR.Using restriction endonuclease and T4 DNA ligase,the CDS fragment was subsequently cloned into pBI121 vector.[Result] The recombinant plasmid was transformed into Agrobacterium GV3101 cells.Wild type plants were then infiltrated using floral-dip method and screened to isolate the desired transgenic lines.[Conclusion] PTR3 over-expression vector was constructed successfully,and transgenic plants were obtained.These works will lay a foundation for further studies.
关键词
拟南芥/PTR3基因/过表达载体/转基因植株Key words
Arabidopsis thaliana/PTR3/Over-expression vector/Transgenic lines引用本文复制引用
出版年
2016
国家科技期刊平台
NSTL
维普
万方数据