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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
引用本文:关继羽,李松励,张鹏,宋光启,刘丽梅,刘颖,王中伟,高飞,唐博,陈育新,岳占碰,李子义.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[J].中国兽医学报,2011,31(4).
作者姓名:关继羽  李松励  张鹏  宋光启  刘丽梅  刘颖  王中伟  高飞  唐博  陈育新  岳占碰  李子义
作者单位: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
基金项目: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。

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

Construction of eukaryotic expression vector pIRES2-EGFP-V13KL and its expression in COS-7 cells and mouse embryos
GUAN Ji-yu,LI Song-li,ZHANG Peng,SONG Guang-qi,LIU Li-mei,LIU Ying,WANG Zhong-wei,GAO Fei,TANG Bo,CHEN Yu-xin,YUE Zhan-peng,LI Zi-yi.Construction of eukaryotic expression vector pIRES2-EGFP-V13KL and its expression in COS-7 cells and mouse embryos[J].Chinese Journal of Veterinary Science,2011,31(4).
Authors:GUAN Ji-yu  LI Song-li  ZHANG Peng  SONG Guang-qi  LIU Li-mei  LIU Ying  WANG Zhong-wei  GAO Fei  TANG Bo  CHEN Yu-xin  YUE Zhan-peng  LI Zi-yi
Institution:GUAN Ji-yu1,LI Song-li1,ZHANG Peng1,SONG Guang-qi1,LIU Li-mei1,LIU Ying1,WANG Zhong-wei1,GAO Fei1,TANG Bo1,CHEN Yu-xin2,YUE Zhan-peng1,LI Zi-yi1*(1.College of Animal Science and Veterinary Medicine,Jilin University,Changchun 130062,China,2.Changchun Protelight Pharmaceutical and Biotechnology Co.,Ltd,Changchun 130012,China)
Abstract: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 sh...
Keywords:V13KL  COS-7 cell  transient expression  flow cytometry  pronuclear injection  
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