炎症性肠病相关基因逆转录病毒表达载体的构建

Construction of retroviral vector carrying human major histocompatibility complex class I chain-related gene A

目的构建炎症性肠病相关基因(major histocompatibility complex class I chain-related gene A,MI-CA)逆转录病毒表达载体pCLXSN-MICA,为进一步研究MICA基因的功能奠定基础。方法根据MICA基因的核苷酸序列,设计并合成引物,通过RT-PCR方法,从含有MICA基因的Hela细胞中,扩增该基因外显子片段,与逆转录病毒表达载体pCLXSN连接,构建MICA基因的逆转录表达载体,并进行鉴定。结果构建的新质粒经PCR、酶切鉴定和DNA测序,证实新质粒构建成功。结论重组MICA逆转录病毒表达载体的构建为进一步研究MICA在炎症性肠病等自身免疫性疾病的作用奠定了基础。

Objective To construct a retroviral vector carrying human major histocompatibility complex class I chain-related gene A （MICA）. Methods MICA eDNA was amplified by RT-PCR from He1a cells. The PCR product was cloned into pCLXSN retroviral vector. The recombined plasmid was identified by PCR method, restriction enzyme reaction and sequence analysis. Results MICA gene was cloned into pCLXSN retroviral vector successfully. Conclusion Retroviral vector carrying MICA eDNA will lay foundation for further studying MICA function in inflammatory bowel disease and other autoimmune diseases.