摘要
目的获取BALB／c小鼠Ii链编码基因,构建真核表达载体并在真核细胞中表达。方法RT-PCR获取BALB／c小鼠Ii链编码基因,插入pEGFP真核表达载体,脂质体转染COS-7细胞,荧光显微镜和激光共聚焦观察外源基因的表达。结果外源基因在COS-7细胞中得到高效表达,激光共聚焦的结果显示,外源基因定位在细胞的内膜系统,并能够和I-Ad分子形成聚集体。结论Ii链在真核细胞中表达后定位在细胞的内膜系统并能和I-Ad分子形成聚集体。
Objective To get Ii chain encoding region, and construct a eukaryotic expression vector, that express efficiently in Mammalia cell line. Methods The encoding gene of BALB/c mouse was obtained by RT-PCR method, then the gene segments were inserted into pEGFP vector. COS-7 cell line was transfected by Li- pofectAMINE2（DO. The expression of foreign gene was demonstrated by laser focus confocal microscope （LFCM）. Results The foreign gene was efficiently expressed in COS-7 cell line. The LFCM results indicated the expressed protein localized in inner membrane system. It could assemble with I-A^d molecular after synthesis. Conclusion Ii encoding gene could express efficiently in Mammalia cell line and assemble with I-A^d molecular after synthesis.
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2006年第6期481-483,共3页
Chinese Journal of Microbiology and Immunology
基金
国家自然科学基金资助项目(30200245)
