Construction Of The Plasmid For Expression Of EGFP-M-IL-2(⁸⁸Arg, ¹²⁵Ala) Fusion Protein And Its Anti-tumor Effects

Objective To construct an eukaryotic expressive vector of p LEGFP-C1-M-IL-2(88Arg,125Ala), p EGFP-N3-M-IL-2(88Arg,125Ala), and to study the expression of this gene in the He La, Glioma cell line U87, and to detect its antitumor activities of recombinant M-IL-2protein. Methods The target fusion gene M-IL-2 was amplified by PCR. The fusion gene was transfected into He La, Glioma cell line U87, by Lipofectamine 2000 immediately after it was confirmed by restrictive enzyme analysis and sequencing. RT-PCR and Western blot were used to confirm expression of the fusion gene in the He La, U87.Prohibitory effect of recombinant M-IL-2 protein on He La, Glioma cell line U87, was assessed by CCK-8 assay and flow cytometer. Results Restrictive enzyme digestion and sequence analysis revealed that M-IL-2 fusion gene was cloned into the eukaryotic expression vector p EGFP-N3, p LEGFP-C1 successfully. Fusion M-IL-2 gene could express in He La, Glioma cell line U87 cells and the fusion protein could inhibit the growth of U87 cells. Conclusion The eukaryotic expression plasmid p EGFP-N3-M-IL-2(88Arg,125Ala), p LEGFP-C1-M-IL-2(88Arg,125Ala) were constructed successfully. Fusion M-IL-2 gene could express in He La, Glioma cell line U87 cells,which laid a foundation for further research of gene therapy.