CryIA(c)和gna融合基因植物表达载体的构建  

Construction of Expression Vector Harboring CryIA(c) and gna Fusion Gene

作　　者：冯翠莲[1,2] 张树珍[1] 

机构地区：[1]中国热带农业科学院热带生物技术研究所,海口571101 [2]海南大学农学院,海南儋州571737

出　　处：《热带作物学报》2010年第2期224-228,共5页Chinese Journal of Tropical Crops

基　　金：中央级公益性科研院所基本科研业务费(No.ITBBZD0724);中国热带农业科学院院基金(No.Rky0617);现代农业产业技术体系建设专项基金(No.nycytx-24)资助

摘　　要：采用重叠延伸PCR技术(gene splicing by overlap extension PCR,简称SOE PCR),通过连接多肽FMDV2A序列将CryIA(c)和gna基因相融合,得到CryIA(c)-2A-gna抗虫融合基因。并进一步将玉米UBI启动子和融合基因(CryIA(c)-2A-gna)分别连接到植物表达载体pCAMBIA3300上,构建成由玉米UBI启动子驱动抗虫融合基因的植物表达载体pNUBG。经过限制性酶切分析鉴定,结果表明,含有融合基因的植物表达载体构建成功。CryIA（c）and gna genes were ligated with FMDV2A by using the technique of gene splicing by overlap extension PCR（SOE PCR）to produce an insect-resistant fusion gene CryIA（c）-2A-gna.Then the maize＇s UBI promoter and the fusion gene were cloned into the plant expression vector pCAMBIA3300 to construct a plant expression vector pNUBG consisting of UBI promoter-driven insect-resistant fusion gene.These recombinant plasmids were confirmed by restriction enzyme and PCR analysis,and the results showed that these recombinant plasmids were constructed successfully.The recombination plasmids were then introduced into Agrobacterium tumefaciens EHA105 by the freeze-thaw transformation method.This work provides a foundation for the transformation of insect-resistant fusion genes to plant mediated by Agrobacterium.

关 键 词：重叠延伸PCR FMDV2A 融合基因 表达载体 

分 类 号：Q78[生物学—分子生物学]