Abstract
CD69 is a type-two transmembrane protein belonging to the NK gene complex family. It is a disulfide-linked surface homodimer with C-type lectin motif and an intracellular signal transduction domain. CD69 is constitutively phosphorylated when expressed on cell memrbane, although the mechanisms of regulating its phosphorylation is unknown. Also, the physiological ligand of CD69 remains unknown. Previous studies had implied that CD69 was involved in HIV and autoimmune disease like lupus and arthritis. Hence, the immunological function of CD69 is of both medical and biological significance. The aims of this study are to clone the tentative CD69 ligand, and study the effect of CD69 on cell proliferation. We have generated CHO-69 cell lines that stably express CD69 on its membrane, and compare it with untransfected CHO cells with cell cycle analysis, and cell number counts. We also make cell lines that stably express CD69-Ig fusion protein, and purified CD69-Ig proteins for FACS screening to identify CD69 binding partners on cell membrane. Lastly, hybridomas that produce anti-CD69 monoclonal antibodies are obtained by immunizing balb/c mice with MBP-CD69 fusion proteins. More experiments are on-going that extends the current report, for example, using CD69-Ig fusion protein to immunoprecipitate its associated proteins from PBMC, and send the CD69 associated proteins for protein sequencing.簡介 Introduction ……………………………………… 3材料與方法 Materials and Methods …………………… 8結果與討論 Results and Discussions…………………… 19參考資料 References ………………………………… 35附錄 Appendix ………………………………………… 39