Construction of the eukaryotic recombinantexpression plasmid of MAGE-3

Objective: To construct MAGE-3 aim gene eukaryotic recombinant expression plasmid pcDNA(3.1)-MAGE-3.Methods: MAGE-3 aim gene containing BamHⅠ,EcoRⅠ enzyme cutting sites was produced from liver cancer tissues by RT-PCR.The aim gene was orientating cloned into pGEM-T Easy vector and subcloned into pcDNA3.1 vector by DNA reconbinant technical.Ampicillin(Amp) antibiotic,blue-white screenexperiment,primer amplify methods were used to selected and identified positiveclone,and the sequence of MAGE-3 aim fragments to be inserted in the middle ofthe position clone was sequenced.Results: MAGE-3 aim gene fragments were amplified.The sequence of MAGE-3 aim gene in the middle of positive recombinant plasmid was compared with the relevant sequence of MAGE-3 of GenBank published after DNA sequenced;and the result was the same completly.Conclusion: The researchsuccessfully constructs eukaryotic recombinant expression plasmid pcDNA3.1-MAGE-3,and this provides the condition for tumor immune treatment.