逆转录病毒载体介导的蜘蛛拖丝蛋白转基因小鼠的制备  

Preparation of retrovirus-mediated spider dragline silk gene transgenic mice

作　　者：杜文敬 梁洋 朴善花 王春生 安铁洙 DU Wenjing;LIANG Yang;PIAO Shanhua;WANG Chunsheng;AN Tiezhu(College of Life Science,Northeast Forestry University,Harbin 150040,China)

机构地区：[1]东北林业大学生命科学学院

出　　处：《黑龙江畜牧兽医》2018年第17期10-12,230,共4页Heilongjiang Animal Science And veterinary Medicine

基　　金：中央高校基本科研业务费专项C类项目(2572016CA10);黑龙江省自然科学基金项目(C2016012)

摘　　要：为了建立通过转基因动物获得蜘蛛拖丝蛋白的方法,试验利用前期构建的蜘蛛拖丝蛋白基因（2S）逆转录病毒载体侵染小鼠胚胎干细胞（embryonic stem cell,ES）,通过嵌合体法制备转基因小鼠,并用PCR技术进行鉴定。结果表明：成功获得嵌合体小鼠;目的基因2S-IRES-EGFP已整合入F0代及F1代嵌合体小鼠的鼠尾基因组中。说明利用逆转录病毒可成功获得表达蜘蛛拖丝蛋白基因（2S）的转基因小鼠。The aim of the present study to establish a method for obtaining spider dragline silk protein from transgenic animals.The mouse embryonic stem cells（ ES） were infected with retroviral stocks carrying spider dragline silk gene（ 2 S）,which were generated in our laboratory previously. The chimeric mice were produced by injection of 2 S-positive ES cells into mouse blastocysts,and identified by PCR technology. The results showed that the chimeric mice were obtained,and the PCR results indicated that the target gene 2 S-IRES-EGFP had been successfully integrated into the tail genome of the F0 and F1 chimeric mice. It was indicated that the transgenic mice expressing spider dragline silk protein gene（ 2 S） could be successfully obtained by retrovirus.

关 键 词：蜘蛛拖丝蛋白基因 逆转录病毒载体 小鼠胚胎干细胞 嵌合体 转基因 显微注射 

分 类 号：S865.1[农业科学—野生动物驯养] Q813.7[农业科学—畜牧兽医]