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[Cloning of human testicular carcinoma antigen MAGE-E1 gene and its expression in E.coli]
1Department of Pathology,Xijing Hospital, Fourth Military Medical University, Xi'an 710032,China.
Aim:
To clone and express the testicular carcinoma antigen MAGE-E1 gene in E.coli.
Methods:
The cDNA encoding human MAGE-E1 gene was amplified by RT-PCR from human glioma cell line BT-325, then the MAGE-E1 gene was inserted into plasmid pGEM-T easy. After sequencing, the MAGE-E1 was cloned into the prokaryotic expression vector pGEX-4T-2 to construct the recombinant expression vector pGEX-4T-2-MAGE-E1 which was used to transform E.coli.
Results:
The expressed product reached the highest level at 5 h after IPTG induction. SDS-PAGE and scanning analysis of gel density indicated that the expressed protein was about M(r) 41 000 and account for 35% of the total bacterial protein.
Conclusion:
The high efficient expression of the MAGE-E1 gene fragment lays the foundation for further preparing antibody against MAGE-E1 protein and the tumor vaccine for glioma immunotherapy.
