[Cloning of human testicular carcinoma antigen MAGE-E1 gene and its expression in E.coli].
OBJECTIVE
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.


CONCLUSIONS
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.
Wang
Li
L
Department of Pathology,Xijing Hospital, Fourth Military Medical University, Xi'an 710032,China.
Li
Qing
Q
Guo
Ai-Lin
AL
Zhang
Feng
F
Guo
Bin
B
Fan
Dai-Ming
DM
D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D005822:Genetic Vectors; D006801:Humans; D010957:Plasmids