经济动物学报2012，Vol.16Issue(1)：26-30,5.
中国林蛙抗菌肽Temporin-1CEa基因的真核表达载体构建。
Construction of Eukaryotic Expression Vector with Rana Antimicrobial Peptides Gene Temporin-lCEa
摘要
Abstract
In order to establish a method to get a large number of antimicrobial peptides from Rana chensinensis,a series of experiments were conducted as follows.According to Chinese frog skin antimicrobial peptides Temporin-1CEa gene mRNA sequence（EU624139） in GenBank,a pair of specific primers were designed and cDNA was obtained from Chinese forest frog skin RNA by reverse transcription.Temporin-1CEa gene coding sequence was amplified using the cDNA,and linked with pEASY-T3 cloning vector.The GFP gene was inserted into the recombinant plasmid Tem-T3 by molecular methods.The Tem-GFP fragment was linked with eukaryotic expression vector pcDNA3.1,and Tem-GFP-pcDNA3.1 recombinant plasmid was achieved finally.Using of lipid infection method,the plasmids were transfected into sheep fibroblast cells,the green fluorescence was observed under a fluorescence microscope after 48 h.qPCR data showed that Tem-GFP fusion protein expression level of transfected Tem-GFP-pcDNA3.1 sheep fibroblasts increased about 300 folds than that of the control group.This study supplied the technical basis for developing mammary gland bioreactor of expressing Temporin-1CEa gene.关键词
中国林蛙/抗菌肽/绵羊成纤维细胞/瞬时表达Key words
Rana chensinensis/antimicrobial peptide/sheep fibroblast cell/transient expression activityanalysis分类
农业科技引用本文复制引用
张志崇,王春生,张秋婷,朴善花,苗向阳,安铁洙..中国林蛙抗菌肽Temporin-1CEa基因的真核表达载体构建。[J].经济动物学报,2012,16(1):26-30,5.基金项目
国家转基因生物新品种培育科技重大专项（2009ZX08008-004B）,国家自然科学基金资助项目 （）
