小鼠釉丛蛋白成熟肽编码区cDNA克隆

Cloning of mouse tuftelin encoding mature protein

目的 :克隆小鼠釉丛蛋白 (tuftelin)成熟肽编码区基因。方法 :设计引物 ,以小鼠牙胚cDNA文库为模板 ,利用PCR方法 ,扩增出小鼠釉丛蛋白成熟区的基因片段 (约 12 0 0bp) ,将所得基因片段插入pBS质粒载体 ,转化到大肠杆菌XL - 1-Blue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 -tuftelin的酶切图谱和序列分析结果与国外文献报道一致。结论 :克隆到小鼠釉丛蛋白成熟肽编码区基因。

AIM: Cloning and partially sequencing of mouse tuftelin encoding mature protein. METHODS: The desired DNA product was obtained from the mouse dental germ cDNA library by PCR with two gene specific primers. The segment (about 1200 bp) was inserted into pBluescript vector, and the interesting plasmid was transformed into E. Coli host strain XL-1-Blue. The double-stranded DNA of the positive clone was analyzed by restriction endonuclease mapping and DNA sequencing. RESULTS: The restriction endonuclease map and sequence of mouse tuftelin encoding mature protein were consistent with those of the references appeared. CONCLUSION: The mouse tuftelin cDNA encoding mature protein was obtained for further study.