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目的：克隆小鼠成釉蛋白（ＡＭＢＮ）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中扩增出小鼠成釉蛋白成熟区的基因片段（约１．１ｋｂｐ），将所得基因片段插入ｐＴＺ１８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，通过限制性酶切和核苷酸序列分析鉴定阳性克隆。结果：重组质粒ｐＴＺ－ＡＭＢＮ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：克隆到小鼠成釉蛋白成熟肽编码区基因
Objective: To clone the coding region gene of mouse mature ameloblastin (AMBN). 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 1.1kbp) was inserted into the pTZ18 plasmid vector and transformed into E. coli DH5α. The positive clones were selected and the recombinant plasmid DNA was extracted. The restriction endonucleases and nucleotides Sequence analysis identified positive clones. Results: The results of restriction enzyme digestion and partial sequence analysis of recombinant plasmid pTZ-AMBN were consistent with those reported in foreign literature. Conclusion: The mouse mature amelogenin coding region gene was cloned