Construction of the Enhanced Yellow Fluorescent Protein Expression Vector Carrying IFN-γ Gene  

作　　者：YuqingLan GeJ 等 

机构地区：[1]SunYat－SenMemorialHospital,SunYat－senUniversityofMedicalScience,Guangzhou510120,China [2]ZhongshanOphtha

出　　处：《眼科学报》2001年第3期154-157,共4页Eye Science

基　　金：This paper is granted by National Nature Science Foudation of China(No.39700153);Natural Science Foundation of Guangdong Province(No.970082)

摘　　要：Purpose: To construct the enhanced yellow fluorescent protein (EYFP) vector carryinginterferon-y gene (ifn-γ) in order to provide an ideal reporter in the expression of ifn-γand location of protein in vitro and in vivo.Method: According to the nucleotide sequence of ifn-y gene, a pair of oligonucleotideswas designed as primer whose two end contained nucleotide sequence of EcoR V and NotⅠ restriction endonuclease respectively. The gene encoding for inf-y was amplified usingPCR technique. After the PCR product was retrieved and purified, it was digested withEcoR V and Not Ⅰ restriction endonuclease, and then cloned into the plasmidpIRES-EYFP. The recombinant plasmid plRES-EYFPIFN-γwas identified by restrictionendonuclease enzyme analysis and DNA sequence analysis.Results: The ifn-γ was successfully amplified and verified by partial DNA sequenceanalysis. The recombinant plasmid was correctly screened.Conclusion: The EYFP expression vector carrying ifn-γgene was successfully established.This research work has formed a base for monitoring the ifn-y gene expression andprotein position in living cells.Purpose: To construct the enhanced yellow fluorescent protein (EYFP) vector carryinginterferon-r gene (ifn-γ) in order to provide an ideal reporter in the expression of ifn-γand location of protein in vitro and in vivo.Method: According to the nucleotide sequence of ifn-γ gene, a pair of oligonucleotideswas designed as primer whose two end contained nucleotide sequence of EcoR V and NotI restriction endonuclease respectively. The gene encoding for inf-r was amplified usingPCR technique. After the PCR product was retrieved and purified, it was digested withEcoR V and Not I restriction endonuclease, and then cloned into the plasmidpIRES-EYFP. The recombinant plasmid pIRES-EYFPIFN-γ was identified by restrictionendonuclease enzyme analysis and DNA sequence analysis.Results: The ifn-γ was successfully amplified and verified by partial DNA sequenceanalysis. The recombinant plasmid was correctly screened.Conclusion: The EYFP expression vector carrying ifn-γ gene was successfully established.This research work has formed a base for monitoring the ifn-r gene expression andprotein position in living cells. Eye Science 2001; 17 : 154 ~ 157.

关 键 词：黄色荧光蛋白质 分子克隆 Γ-干扰素基因 

分 类 号：Q785[生物学—分子生物学]