棉花脂肪酸脱氢酶-2(FAD2)转基因小鼠模型的建立  

Establishment of Transgenic Mice Integrated with Gene Encoding Cotton Omega-6 Fatty Acid Desaturase-2

作　　者：王宇[1] 安晓荣[1] 柳青 苟克勉[1] 

机构地区：[1]中国农业大学生物学院农业生物技术国家重点实验室,北京100094 [2]BlackmountainLaboratory,CSIROPlantIndustry,Canberra,Australia

出　　处：《农业生物技术学报》2005年第2期207-211,共5页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金(No.30270647);和北京市自然科学基金项目(No.5012007)资助

摘　　要：以pEGFP-N1质粒为基础载体,切除gfp基因的编码序列后,在多克隆位点依次插入动物肌肉α-actin启动子和棉花ω-6脂肪酸脱氢酶编码基因fad2(fattyaciddesaturase-2)的cDNA序列,构建成fad2cDNA真核表达载体。用显微注射方法,将线性化的载体DNA注入KM×B6D2F1小鼠受精卵的的原核内部,然后将胚胎移植到输卵管内,13只受体怀孕产仔。合计移植了286枚受精卵,最后获得34只健康的成年小鼠,PCR方法检测确认5只为阳性小鼠,Southernblot证明只有1只阳性转基因小鼠。The aim of this study is to produce the transgenic mouse integrated with gene encoding cotton (Gossypium hisrutum) omega-6 fatty acid desaturase-2 (FAD2). We firstly constructed an expression vector bearing the fad2 cDNA derived by α-actin promoter of bovine skeletal muscle gene. Then the linear vector was microinjected into the pronucleus of the KM×B6D2F1 zygotes to produce the transgenic mice. Thirty-four adult mice were produced from 286 of injected embryos. Five of them were transgenic by genomic DNA polymerase chain reaction (PCR). Southern blotting results revealed that only one mouse was integrated with fad2 gene.

关 键 词：转基因小鼠模型 脱氢酶 脂肪酸 棉花 Southern cDNA序列 真核表达载体 GFP基因 多克隆位点 PCR方法 编码序列 编码基因 动物肌肉 acid 注射方法 胚胎移植 成年小鼠 blot 受精卵 启动子 线性化 阳性 质粒 原核 受体 

分 类 号：Q78[生物学—分子生物学] X703[环境科学与工程—环境工程]