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

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.