真核表达载体pIRES2-EGFP-V13KL的构建及在COS-7细胞和小鼠胚胎中的表达  

Construction of eukaryotic expression vector pIRES2-EGFP-V13KL and its expression in COS-7 cells and mouse embryos

作　　者：关继羽[1] 李松励[1] 张鹏[1] 宋光启[1] 刘丽梅[1] 刘颖[1] 王中伟[1] 高飞[1] 唐博[1] 陈育新 岳占碰[1] 李子义[1] 

机构地区：[1]吉林大学畜牧兽医学院动物胚胎工程吉林省重点实验室,吉林长春130062 [2]长春普莱医药生物技术有限公司,吉林长春130012

出　　处：《中国兽医学报》2011年第4期548-552,共5页Chinese Journal of Veterinary Science

基　　金：国家转基因生物新品种培育重大专项(2008ZX08008-005);国务院华侨办华人创业团队基金重点专项资助项目

摘　　要：构建了真核表达载体pIRES2-EGFP-V13KL,瞬时转染COS-7细胞及对小鼠受精卵进行原核注射,旨在通过抗菌肽V13KL在COS-7细胞、小鼠胚胎中的表达情况评价其对细胞生理状态、胚胎发育方面可能产生的影响。结果显示,V13KL可以与EGFP在COS-7细胞中高效共表达,转染效率为75.8%,且原核注射后约10%的胚胎发育成带有绿色荧光的囊胚。The V13KL is a kind of antimicrobial peptides through optimal design and artificial synthesis,which it is characteristic of stronger antimicrobial activity and weaker hemolytic activity.This study was conducted to examine expression of this peptide in COS-7 cells and detect the effect of this peptide on early development of mouse embryos.The eukaryotic expression vector pIRES2-EGFP-V13KL was constructed,and transfected into the COS-7 cells and injected into the pronucleus of the mouse embryos.The results showed that V13KL and EGFP could co-express in COS-7 cells,and the transfection efficiency is 75.8%.About 10% of the mouse embryos injected with the vector developed into blastula.This study may be the basis of large-scale application and the cultivation of transgenic animals in the future.

关 键 词：V13KL COS-7 cell 瞬时表达 流式细胞术 原核注射 

分 类 号：Q78[生物学—分子生物学] S852.1[农业科学—基础兽医学]