BALB/c小鼠I-A^dαβ链和恒定链Ii在COS-7中的共定位

Co-localization of BALB/c mice I-A^dαβ chains and invariant chain in COS-7 cell line

目的 :研究BALB/c小鼠I Adαβ链和恒定链Ii分子p4 1在COS 7细胞中的定位 ,探索抗原提呈的分子规律。方法 :构建了带有红色荧光蛋白标签的I Adαβ链真核双顺反子表达载体 pRed IRES I Ad和带有绿色荧光蛋白标签的恒定链真核表达载体 pEGFP Ii,使用Lipofectamine 2 0 0 0转染COS 7细胞 ,用激光共聚焦显微镜观察 2个外源蛋白在细胞中的共定位。结果 :I Adαβ分子在COS 7细胞中能够形成聚集状态 ,并且和Ii链共同定位于细胞的内膜系统。结论 :小鼠mIip4 1能够和I Adαβ分子在COS 7细胞中形成聚集体结构 ,mIip4 1分子的导向肽并不具有选择性导向作用 ,聚集体向细胞膜表面的移动很可能是通过胞吐的方式进行。

Objective: To investigate the co-localization of BALB/c mice I-A d αβ chains and invariant chain in COS-7 cell line. Methods: pRed-IRES-I-A d and pEGFP-Ii were constructed respectively. COS-7 cell line was transfected with the plasmids by using Lipofectamine 2000. The expression of foreign genes were detected by laser confocus microscope. Results:Results of laser confocus microscopy showed the subcellular localization of target proteins. The results indicated that mIip41 was over synthesized in ER. Some mIip41 could form nonomeric complexes with αβ heterodimers, but some could not. In opposition to other research groups,the N terminus of mIip41 was not found to have the specific characteristics in endosomal targeting. Conclusion:mIip41 can form nonomeric complexes with αβ heterodimers. The cytoplasmic domain of mIip41 do not have the specific characteristics in endosomal targeting.