摘要
目的 :从人牙乳头细胞内克隆骨形成蛋白 (BMP)细胞内信号转导基因 Smad1的功能性结构域— MH2结构域。方法 :原代培养人牙乳头细胞 ,从培养的细胞中提取总 RNA,逆转录合成 c DNA第 1条链 ;设计上下游引物 ,进行 RT- PCR,扩增 Smad1基因 MH2结构域的基因片段 ;将所获得的基因片段定向插入 PUC19载体 ;转化大肠杆菌 JM10 9,挑选阳性克隆 ,鉴定后进行序列测定。结果 :获得的 Smad1MH2 结构域的 c DNA片段大小为 44 4bp,并且成功构建 PU C19/ MH2重组质粒。结论 :人牙乳头细胞内存在 Smad1信号转导途径 ,BMP调控人牙乳头细胞分化可能是通过
Objective:To clone and sequence cDNA of MH2 domain of Smad1 gene from human dental papilla cells. Methods:Total RNA was isolated from primarily cultured human dental papilla cells and reversely transcribed into single stranded cDNA.The desired DNA product was obtained by PCR with two gene specific primers.The segment was inserted into PUC19 vector and the plasmid was transformed into E.coli JM109.The double stranded cDNA of positive clone was sequenced.Results:The sequence of MHz domain of Smad1 cloned from human dental papilla cells was consistent with that reported by Hoodless et al. Conclusion:Smad1 exists in human dental papilla cells,BMP signaling may be mediated by smad1 in human dental papilla cells.
出处
《实用口腔医学杂志》
CAS
CSCD
北大核心
2001年第6期498-501,共4页
Journal of Practical Stomatology
基金
国家自然科学基金资助项目 (39870 789)
