MHCⅡ类反式激活因子基因的克隆、鉴定及初步功能测定

Cloning, identification and preliminary function test of CⅡTA gene

目的 克隆MHCⅡ类反式激活因子 (classⅡtransactivator,CⅡTA)基因并进行初步功能测试 ,为CⅡTA的应用研究奠定基础。方法 RT PCR扩增CⅡTA基因 ,将其克隆到pGEM T载体上并进行酶切和测序鉴定 ;用EcoRⅠ、XhoⅠ将CⅡTA定向克隆到表达载体pcDNA3上 ,用脂质体转染法将pcDNA3/CⅡTA转入HeLa细胞 ;流式细胞术观察细胞表面HLA DR/DQ的表达变化。结果 成功克隆CⅡTA基因 ,CⅡTA基因的转入使HeLa细胞表面表达HLA DR/DQ分子。结论 转入CⅡTA能使HeLa细胞表面表达MHCⅡ类分子表达 ,CⅡTA参与调控MHCⅡ类基因的转录和表达。

Objective Molecular cloning of CⅡTA (classⅡ transactivator) gene and analysis of its function preliminary. Methods CⅡTA gene obtained by RT-PCR was cloned into pGEM-T/pcDNA3 plasmid. Restriction endonuclease and sequence analysis were used to identify the CⅡTA gene. Flow cytometry was used to test the expression of HLA-DR/DQ in HeLa cells transfected by pcDNA3/CⅡTA construction. Results CⅡTA gene clone was obtained and the expression of HLA-DR/DQ in HeLa cells was induced by transfection of CⅡTA gene. Conclusion Transfection of CⅡTA enhanced the expression of MHC class Ⅱ molecules. CⅡTA may play an important role in the transcription and expression of MHC class Ⅱgenes.