转基因香石竹中F3’5’H基因的克隆、表达和免疫学鉴定  

Cloning,Expression and Immunological Identification of F3'5'H Gene in Transgenic Carnation (Dianthus canpphyllus Linn.)

作　　者：白蓝[1,2] 贾军伟[1] 孙建萍[1] 李鹏[1] 赵明文[2] 潘爱虎[1] 

机构地区：[1]上海市农业科学院生物技术研究所,上海市农业遗传育种重点实验室,农业部转基因植物环境安全监督检验测试中心(上海),上海201106 [2]南京农业大学生命科学学院,农业部农业环境微生物工程重点开放实验室,南京210095

出　　处：《植物生理学通讯》2010年第2期108-112,共5页Plant Physiology Communications

基　　金：上海市科技兴农重点攻关项目(沪农科攻字2008第8-9号)

摘　　要：在研究转基因香石竹品系月之霓裳(Moonshade)、月之伊人(Moonlite)中外源基因F3’5’H的表达中,本文克隆了F3’5’H全长基因1.5kb,构建获得工程菌株Escherichia coli BL21(DE3)(+F3'5'H)。SDS-PAGE分析的结果显示,该菌株高效表达出F3’5’H重组蛋白,约占菌体总蛋白的30%。用经纯化的F3’5’H重组蛋白作为抗原,制备F3’5’H重组蛋白的抗血清,经ELISA免疫学分析表明,该抗血清的效价为1:25600。Western blot结果表明F3’5’H重组蛋白具有良好的IgG结合活性,且抗血清与转基因香石竹品系月之霓裳和月之伊人中的外源基因F3’5’H所表达的蛋白发生明显的抗原抗体反应。这样,月之霓裳和月之伊人用于评价转基因香石竹品系的环境安全性在我国也得到了验证。In order to study the expression of exogenous gene F3＇5＇H in transgenic carnation lines Moonshade and Moonlite,the full-length F3＇5＇H gene of 1.5 kb was cloned,and the strain Escherichia coli BL21（DE3） （＋F3＇5＇H） was obtained.SDS-PAGE analysis showed that the high-level expressed recombinant protein F3＇5＇H was detected and the expression level was almost 30% of the total protein.The recombinant protein was used as antigen to obtain the antiserum.And the immunological analysis of ELISA showed the antiserum titer was 1：25 600.Western blot analysis indicated that the recombinant protein F3＇5＇H could combine with sera IgG,and the antiserum showed specific immunological response to the F3＇5＇H protein extracted from Moonshade and Moonlite.

关 键 词：转基因香石竹 F3’5’H 原核表达 免疫学鉴定 

分 类 号：S682.19[农业科学—观赏园艺]