Cloning of human preferentially expressed antigen of melanoma gene and its expression in E.coli
- VernacularTitle:人黑色素瘤特异性抗原基因的克隆及其在原核系统中的表达
- Author:
Li WANG
;
Ailin GUO
;
Shukui QIN
;
Qing LI
;
Li WANG
- Publication Type:Journal Article
- Keywords:
Preferentially expressed antigen of melanoma gene;
Leukemia;
Minimal residual disease;
Gene cloning;
Expression
- From:
Journal of Medical Postgraduates
2003;0(04):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To clone and express the preferentially expressed antigen of melanoma (PRAME) gene in E.coli. Methods: The cDNA encoding human PRAME gene was extracted from human AML cell line HL-60 and amplified by RT-PCR, then the PRAME gene was inserted into plasmid PGEM-T-easy. After sequenced, it was cloned into the prokaryotic expression vector pGEX-4T-2 to construct a clone with high level expression and the recombinant plasmid was cloned in E.coli. Results:The protein product reached the highest level at 5 h after IPTG induction. Conclusion: Since PRAME is only expressed at low levels in a few normal tissues and encodes an antigen recognized by autologous cytotoxic T lymphocytes, while expressed at high levels in patients with leukemia, it might be a new targeting candidate for tumor immunotherapy.
