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

Purpose:To construct the enhanced yellow fluorescent protein (EYFP) vector carrying interferon-γ 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 nocleotide sequence of ifn-γ gene,a pair of oligonucleotides was designed as primer whose two end contained nucleotide sequence of EcoR V and Not I restriction endonuclease respectively .The gene encoding for ,inf-γ was amplified using PCR technique After the PCR product was retrieved and purified,itwas digested with EcoR V and Not I restiction endonuclease,and then cloned into the plasmid p IRES-EYFP.The recombinant plasmid p IRES-EYFPIFN-γ was identified by restriction endonuclease enzyme analysis and DNA sequence analysis .Results:The ifn-γ was successfully amplified and verified by partial DNA seuqence analysis.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-γ gene expression and protein positon in living cells.Eye Science 2001;17:154-157.