南方农业学报2012，Vol.43Issue(9)：1262-1268,7.DOI:10.3969/j:issn.2095-1191.2012.09.1262
高山离子芥冷诱导基因转化甘蔗二元植物表达载体构建
Construction of bivalent plant expression vector with cold-induced gene of Chorispora bungeana in sugarcane
摘要
Abstract
[Objective]A new bivalent plant expression vector, named pCambial300-cbcorl5a-bar, was recombined by inserting two genes, I.e. Cbcor15a and eGFP, into the vector pCambial300-bar and replacing the promoter CaMV 35s with Ubi-1. [Method]Based on the multiple cloning sites of the expression vector pCambial300-bar, the primers were designed according to the nucleotide sequence of gene Cbcorl5a and eGFP and the promoter Ubi-1, and then the fragments of the genes and promoter were inserted into the vector pCambial300-bar. The plasmids of the vector pCambial300-cb-cor15a-bar were transduced into onion epidermal cells via particle bombardment. The results were observed through fluorescence microscopy and confocal laser scanning microscope, respectively. [Result]Compared to onion epidermal cell with pCambia 1300-bar, onion epidermal cell transduced with the recombinant vector plastnid had a very bright green florescence. [Conclusion]Downstream cold-induced gene Cbcorl5a and reporter gene eGFP regulaled by up-stream promoter Ubi-1 were expressed efficiently, which could ensure the construction of Cbcorl5a.关键词
Cbcor15a/甘蔗/转基因/植物表达载体构建/瞬时表达Key words
CbcorlSa/ sugarcane/ transgene/ construction of plant expression vector/ transient expression分类
农业科技引用本文复制引用
卢双楠,刘晓静,李鸣,梁俊,李容柏,李粲,滕峥,刘开雨,刘芳,邱永福,梁朝旭,方位宽,何姗珊..高山离子芥冷诱导基因转化甘蔗二元植物表达载体构建[J].南方农业学报,2012,43(9):1262-1268,7.基金项目
广西科学基金项目(桂科自0990182,桂科基0778006-4,桂科青0832059,2011GXNSFF018002,2011GXNSFD018021) （桂科自0990182,桂科基0778006-4,桂科青0832059,2011GXNSFF018002,2011GXNSFD018021）
广西农业科学院博士后基金项目(桂农科博2009013) （桂农科博2009013）
广西农业科学院基本科研业务专项项目(201107Z基,G2010003,G2010003) （201107Z基,G2010003,G2010003）
广西农业科学院公益性维持项目(桂农科2012GW13) （桂农科2012GW13）
南宁市科学研究与技术开发计划项目(201102026B) （201102026B）
