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目的：克隆小鼠釉原蛋白（ＡＭＧ）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中扩增出小鼠釉原蛋白成熟区的基因片段（约６７０ｂｐ），将所得基因片段插入ｐＢＳ质粒载体，转化到大肠杆菌ＤＨ５α后挑选阳性克隆，提取重组质粒ＤＮＡ，通过限制性酶切和部分核苷酸序列分析鉴定阳性克隆。结果：重组质粒ｐＢＳ－ＡＭＧ的酶切图谱和序列分析结果与国外文献报道一致。结论：克隆到小鼠釉原蛋白成熟肽编码区基因。
OBJECTIVE: To clone the gene coding for the mature peptide of mouse amelogenin (AMG). 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 The gene fragment of mouse amelogenin mature region (about 670 bp) was inserted into the pBS plasmid vector and transformed into E. coli DH5α. The positive clones were selected and the recombinant plasmid DNA was extracted. The recombinant plasmid DNA was digested by restriction enzyme and partial nucleotide sequence Analysis identified positive clones. Results: Restriction map and sequence analysis of the recombinant plasmid pBS-AMG were consistent with those reported in foreign literature. Conclusion: The coding region of murine amelogenin mature peptide was cloned.