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	转杂色鲍抗冻肽-Ⅲ基因表达载体pIRES2-EGFP-AFP-HdiActinP的构建; The Construction of Expression Vector pIRES2-EGFP-HdiActinP-AFP for Transgenic Small Abalone Haliotis diversicolor Using Antifreeze Peptide Ⅲ
	李卫东 ; 陈维宇 ; 柯才焕
	2009
关键词	杂色鲍 抗冻肽 启动子 载体构建 Haliotis diversicolor antifreeze peptide promoter construction vector
英文摘要	现代转基因技术是一种快速改良品种特性的有效方法.采用转基因技术提高我国南方重要贝类经济养殖品种——杂色鲍的耐低温特性是扩大其养殖范围的重要手段.以PIrES2-EgfP为基本载体,以合成并经过功能鉴定的抗冻肽(AfP)和杂色鲍肌动蛋白启动子(HdIACTInP)为插入元件,首先用限制性内切酶bAMHⅠ和ECOrⅠ对基本载体PIrES2-EgfP和AfP-Ⅲ进行双酶切,目的酶切产物被回收后再连接得到载体PIrES2-EgfP-AfP.随后利用限制性内切酶SACⅠ和ECOrⅠ对载体PIrES2-EgfP-AfP和HdIACTInP进行双酶切,目的酶切产物被回收后再连接最终得到杂色鲍转AfP-Ⅲ的表达载体PIrES2-EgfP-AfP-HdIACTInP,经过聚合酶链式反应和测序验证证明载体构建成功.; Small abalone,Haliotis diversicolor is one of important maricultural species which distributes in South China.Modern transgenic technique is a rapid and direct way to improve characterization of aquaculture species.It enables us to breed a new line of small abalone which has a low temperature-resistant feature so that it can be cultured in wider areas than now.In this study,pIRES2-EGFP was used as the basic vector,and the antifreeze peptide Ⅲ and HdiactinP were used as inserting elements.Firstly the vector pIRES2-EGFP and segment AFP were digested with the restriction endonuclease BamHⅠ and EcoRⅠ.The target components of the digested product were ligated consequently and the vector pIRES2-EGFP-AFP was constructed.Secondly the vector pIRES2-EGFP-AFP and segment HdiactinP were digested with restriction endonuclease SacⅠand EcoRⅠ.Then the target components of the digestion product were ligated.Finally the transgenic H.diversicolor antifreeze peptide expression vector pIRES2-EGFP-AFP-HdiActinP was constructed,which was confirmed by polymerase chain reaction(PCR)and sequencing.In summary,our result provided a potential basis for the consequent development of the transgenic H.diversicolor researches.; 国家863计划课题(2006AA10A407);农业科研专项(nyzx07-047);现代农业产业技术体系建设专项(nycytx-47)资助
语种	zh_CN
内容类型	期刊论文
源URL	[http://dspace.xmu.edu.cn/handle/2288/104245]
专题	海洋环境－已发表论文
推荐引用方式 GB/T 7714	李卫东,陈维宇,柯才焕. 转杂色鲍抗冻肽-Ⅲ基因表达载体pIRES2-EGFP-AFP-HdiActinP的构建, The Construction of Expression Vector pIRES2-EGFP-HdiActinP-AFP for Transgenic Small Abalone Haliotis diversicolor Using Antifreeze Peptide Ⅲ[J],2009.
APA	李卫东,陈维宇,&柯才焕.(2009).转杂色鲍抗冻肽-Ⅲ基因表达载体pIRES2-EGFP-AFP-HdiActinP的构建..
MLA	李卫东,et al."转杂色鲍抗冻肽-Ⅲ基因表达载体pIRES2-EGFP-AFP-HdiActinP的构建".(2009).

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