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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 To clone the class Ⅱ transactivator (CⅡTA) gene and carry out preliminary functional tests to lay a foundation for the application of C Ⅱ TA. Methods The CⅡTA gene was amplified by RT-PCR and cloned into pGEM T vector. The recombinant plasmid was digested with restriction endonucleases and sequenced. EcoRⅠ and XhoⅠ were used to clone CⅡTA into pcDNA3. The recombinant plasmid pcDNA3 / Into HeLa cells; the expression of HLA DR / DQ on the cell surface was observed by flow cytometry. Results The CⅡTA gene was cloned successfully. The transfection of CⅡTA gene resulted in the expression of HLA DR / DQ on the surface of HeLa cells. Conclusion Transfection into CⅡTA can induce the expression of MHC class II molecules on the surface of HeLa cells. CⅡTA is involved in the regulation of transcription and expression of MHC class Ⅱ genes.