小鼠牙本质磷蛋白cDNA的克隆和部分序列分析

Cloning and partially sequencing of mouse dentin phosphoprotein cDNA

目的：克隆小鼠牙本质磷蛋白（ＤＰＰ）ｃＤＮＡ。方法：用异硫氰酸胍一步法从小鼠牙胚组织中抽提总ＲＮＡ，用ｏｌｉｇｏ（ｄｔ） 作引物逆转录合成牙胚ｃＤＮＡ，然后利用ＰＣＲ 方法，从ｃＤＮＡ 中扩增出小鼠ＤＰＰｃＤＮＡ基因片段（ 约１ ．６ｋｂ），将所得基因片段插入ｐＢｌｕｅｓｃｒｉｐｔ 质粒载体，转化到大肠杆菌ＸＬ１ － Ｂｌｕｅ 后挑选阳性克隆，提取重组质粒ＤＮＡ，通过酶切分析和核苷酸序列分析鉴定阳性克隆。结果：酶切图谱和部分序列分析结果与国外文献报道一致。结论：克隆到小鼠ＤＰＰｃＤＮＡ 基因片段。

Aim:To clone and partially sequencing mouse dentin phosphoprotein cDNA. Methods: In the study, total RNA was extracted from the mouse tooth germs by acid guanidinium thiocyanata-phenol-chloroform method, the desired DNA product was obtained from the total RNA by RT-PCR with the primers including oligo(dt) and two gene specific primers. The segment(about 1.6kb) was inserted into pBluescript vector and the interesting plasmid was transformed into E. Coli host strain XL1-Blue. The double-stranded DNA of the positive clone was analyzed by restriction endoncuclease mappping and DNA sequencing. Results: The restriction endonuclease map and sequence of mouse DPP cNDA in this study were consistent with those in the references published. Conclusons: The mouse DPP cDNA was obtained for further research.