引用本文
  • 尹国,李世云,路正营,韩永亮,王晔,张彦波,洪伟东,商海红,张朝军,张雪妍,李俊玲,李付广.植物双元表达载体pCRI1210的构建及其功能验证[J].植物遗传资源学报,2017,18(5):960-967.    [点击复制]
  • yinguo,李世云,路正营,韩永亮,王晔,张彦波,洪伟东,商海红,张朝军,张雪妍,李俊玲,李付广.Construction and Function Analysis of a plant binary expression vector pCRI1210[J].植物遗传资源学报,2017,18(5):960-967.   [点击复制]
【打印本页】 【在线阅读全文】【下载PDF全文】 查看/发表评论下载PDF阅读器关闭

←前一篇|后一篇→

过刊浏览    高级检索

本文已被:浏览 71次   下载 39 本文二维码信息
码上扫一扫!
植物双元表达载体pCRI1210的构建及其功能验证
尹国,李世云,路正营,韩永亮,王晔,张彦波,洪伟东,商海红,张朝军,张雪妍,李俊玲,李付广
0
(邯郸市农业科学院;中国农业科学院棉花研究所)
摘要:
本研究在pBI121载体的基础上,在其骨架中插入含有CaMV35S启动子、棉花β-tubulin 基因内含子和CaMV 35S polyA终止子并含有GFP作为报告基因的干涉表达框。通过验证,所插入的干涉表达框能够正常表达插入的外源基因,且GFP基因可以正常表达。该载体被命名为pCRI1210,它是一种双元植物表达载体,既可以用来当做表达载体,又可以用作干涉载体。该载体在TUB intron的5’端和3’端分别具有5个和4个酶切位点,可方便的进行目的基因片段的正反向插入,构建可转录出发卡结构的dsRNA。同时在CaMV 35S启动子的两端都含有多克隆位点,可以方便进行启动子的替换。本研究为转基因研究提供了一种非常实用的工具。
关键词:  载体,pCRI1210,干涉
DOI:10.13430/j.cnki.jpgr.2017.05.019
投稿时间:2017-01-20修订日期:2017-05-15
基金项目:黄淮棉区转基因特早熟棉花新材料及新品种创制(2015ZX08005002-007);国家棉花产业技术体系冀南综合试验站(CARS-18-28);短季棉种质资源与育种技术创新(16226307D)
Construction and Function Analysis of a plant binary expression vector pCRI1210
yinguo,李世云,路正营,韩永亮,王晔,张彦波,洪伟东,商海红,张朝军,张雪妍,李俊玲,李付广
()
Abstract:
In the study, a plant binary expression vector, named pCRI1210, was constructed on the basis of pBI121 vector, which contained CaMV 35S promoter, cotton β-tubulin gene introns , CaMV 35S polyA terminator and interference expression box with GFP as reporter gene. Functional analysis in tobacco showed that the interference express cassette can normally expressed exogenous gene and GFP gene . The vector had multiple cleavage sites in a TUB intron 5 "and 3" , so that target gene can be positive and reverse inserted and hairpin structure of dsRNA was builded. In addition, the vector carries multiple cloning sites at both 5′and 3′of the CaMV 35S promoter, which allows easy exchange 35S promoter to study other promoter functions. pCRI1210 can used to expression vector or interference vector,which provided a very practical tool for transgenic research.
Key words:  carrier, pCRI1210, interference

用微信扫一扫

用微信扫一扫