女性在月經開始時，子宮內膜的腺體會呈現不同型態，目前知道這種不同型態的表現是因為細胞受到荷爾蒙刺激(如 Estrogen 和 Progesterone)造成的。但是相關於其他的基因或蛋白參與此調控機轉的報告，則非常少。之前本實驗室以 DDRT-PCR 鑑定出在增殖期早期(月經週期第五天)和分泌期(月經週期第十八天)的腺體細胞中，有數個表現差異性的基因，如Scramblase、NUDT9、NUDT10、XM_027541等。其中XM_027541為一推測性基因，目前尚未有文獻發表，可能在子宮內膜的週期變化中具有重要的功能。因此我們首先以DE 52、High Q 陰離子交換樹脂等方法將XP_027541 純化出來，然後對蛋白進行定序。此外，利用 Ni-NTA純化Recombinant protein 免疫兔子得到抗體對子宮內膜組織不同時期作 Western blotting 分析，結果發現XP_027541在分泌期偵測到明顯的訊號。而我們從生物資訊的比對發現在NCBI中XM_027541與ATPase class VI type 11A在 Genome 序列中的距離很近只有457的base pair; 但在EBI資料庫中則指出 XM_027541 和ATPase class VI type 11A exon30 會發生重疊。最後藉由RT-PCR 及DNA 定序證明 XM_027541 是存於ATPase exon30 的位置，並推測可能有 Alternative splicing 的機制在其中。 The glandular epithelial cells of the endometrium existed distinct morphology in the menstrual cycle. These differentiates are from the stimulation with many hormone. In our laboratory, we have identified the specific gene expressing in the early proliferative phase or in the secretory phase such as phospholipids scramblase, NUDT9, and XP_027541. We focus on the XP_027541 because it may exist some important function in regulating the rule change of endometrium. We first purified the 13 kDa protein and sequencing. And then we get the recombinant protein XP_157-114 and XP_2115-217, and immune the rabbit to gain the polyclonal antibody. We found XP_027541 just near 457 base pair to the ATPase classVI type 11A. But in EBI data base, we found XM_027541 were over lapping with the ATPase class VI type 11A exon 30 region. At last we identified that XM_027541 were located in the exon 30 of ATPase class VI type 11A by PCR and sequencing to suggest that ATPase class VI type 11A may exist the mechanism of RNA alternative slicing.
