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目的：克隆小鼠牙本质涎蛋白（ＤＳＰ）成熟肽编码区基因。方法：用异硫氰酸胍一步法从昆明新生小鼠磨牙牙胚组织中抽提总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ＤＳＰ成熟区的基因片段（约１．４ｋｂｐ），将所得基因片段插入ｐＢＳ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，通过限制性酶切和核苷酸序列分析鉴定阳性克隆。结果：重组质粒ｐＢＳ－ＤＳＰ的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ＤＳＰ成熟肽编码区基因，正在进行该基因的表达和活性鉴定。
OBJECTIVE: To clone the coding region of mouse mature protein of dentin sialoprotein (DSP). METHODS: Total RNA was extracted from tooth germ tissue of Kunming newborn mice by one-step guanidine isothiocyanate method. Oligo (dt) was used as a primer to reversely synthesize the tooth germ cDNA. PCR was then used to amplify the cDNA from the cDNA Mouse DSP mature region of the gene fragment (about 1.4kbp), the resulting gene fragment was inserted into the pBS plasmid vector, transformed into E. coli XL1-Blue positive clones after selection, the recombinant plasmid DNA was extracted by restriction enzyme and nucleotide Sequence analysis identified positive clones. Results: Restriction map and partial sequence analysis of recombinant plasmid pBS-DSP were consistent with those reported in foreign literature. Conclusion: The coding region of mature peptide of mouse DSP was cloned and the gene expression and activity identification were underway.