AIM: To clone and sequence the gene encodin g hBSP mature peptide from human dental pulp cells. METHOD: Total RNA was isolated from cultured human dental pulp cells and desired cDNA fragment was obtained by RT-PCR with two gene specific primers. The segment was inserrted into pBluescript KS vector and the result plasmid was ransformed int o DH5α. The positive clone and sequence were performed. RESULTS: The 900bp specific fragment was obtained. The sequence of the gene encodi ng hBSP mature pepide coincides with that of the references published. CONCLUSION: The gene encoding hBSP mature peptide was obtained from human dental pulp cells. This result will help us to further investigate the expression and function of hBSP.%目的:证实人牙髓细胞中骨涎蛋白(Bone sialopro tein, BSP)的表达,获得BSP成熟肽基因。方法:体外培养人牙髓 细胞,提取人牙髓细胞总RNA,反转录合成cDNA,以特异性引物扩增BSP成熟肽基因并克隆到 pBluescript KS载体,筛选阳性克隆进行序列测定。结果:PCR扩增 到一特异性的约900bp的片段,克隆后筛选出阳性克隆进行序列分析,测定结果表明与国外 已发表的序列完全一致。结论:人牙髓细胞中有BSP基因的表达,并 克隆得到BSP成熟肽基因,为表达有活性的BSP蛋白,进一步研究其生物学功能奠定基础。
展开▼