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Purpose: To construct the enhanced yellow fluorescent protein (EYFP) vector carryinginterferon-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 nucleotide sequence of ifn-r 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-γ 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.Thi
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 nucleotide sequence of ifn-r gene, a pair of oligonucleotides was designed as primer whose two end contained nucleotide sequence of EcoR V and NotI restriction endonuclease respectively. The gene encoding for inf-γ was amplified using PCR technique. After the PCR product was retrieved and purified , it was digested with EcoR V and Not I restriction endonuclease, and then cloned into the plasmid pIRES-EYFP. The recombinant plasmid pIRES-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 sequenceanalysis. The recombinant plasmid was correctly screened. Conlusion: The EYFP expression vector carrying ifn-γ gene was successfu lly established.Thi