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目的：克隆人牙本质基质蛋白１（ＤＭＰ１）成熟肽编码区基因片段。方法：用异硫氰酸胍一步法从引产胎儿牙乳头组织中抽提总ＲＮＡ，用Ｏｌｉｇｏ（ｄｔ）作引物逆转录合成牙乳头ｃＤＮＡ，然后利用ＰＣＲ方法，从ｃＤＮＡ中扩增出人ＤＭＰ１成熟区的基因片段（约１．８ｋｂｐ），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：克隆到人ＤＭＰ１成熟肽编码区基因片段。
Objective: To clone the gene fragment encoding human dentin matrix protein 1 (DMP1) mature peptide. METHODS: Total RNA was extracted from fetal papilla tissue by using one-step guanidinium thiocyanate method and reverse transcription of Oligo (dt) primer was used to synthesize dental papilla cDNA. Then PCR amplification was used to amplify human DMP1 from cDNA Region of the gene fragment (about 1.8kbp), the resulting gene fragment inserted into the pBluescript plasmid vector, transformed into E. coli XL1-Blue positive clones after selection, the recombinant plasmid DNA was extracted by restriction analysis and nucleotide sequence analysis identified positive clones . Results: The results of enzyme digestion and partial sequence analysis were consistent with those reported in foreign literature. Conclusion: The human DMP1 mature peptide coding region gene fragment was cloned.