小鼠釉丛蛋白基因不同mRNA剪接体的克隆

Cloning of the different transcripts of mouse tuftelin mRNA

目的 :克隆小鼠釉丛蛋白基因不同mRNA剪接体 ,分析小鼠釉丛蛋白的多态性。方法 :采用异硫酸氰胍一步法从新生小鼠磨牙牙胚组织中提取总RNA ,反转录成cDNA第一链 ,经PCR扩增出小鼠釉丛蛋白基因特异片段 ,插入pBS质粒 ,筛选阳性克隆 ,并经酶切和核苷酸序列分析鉴定阳性克隆。结果 :克隆T1序列与已发表的完整釉丛蛋白mRNA序列一致。克隆T2序列与完整釉丛蛋白mRNA序列比较 ,缺失序列的 84～15 8的 75个碱基片段。结论 :小鼠釉丛蛋白基因在转录mRNA时 ,可产生不同的剪接体。

AIM: It is reported that there are different forms of tuftelin protein. The aim of this study is to clone the different transcripts of mouse tuftelin. METHODS: The total RNA was extracted from the tooth germs of newborn mouse by acid guanidinium isothiocyanate-phenol-chloform method. Then the first strand of cDNA was transcriptased with the total RNA as model,and the desired cDNA segments was obtained from the first strand cDNA by PCR.The retracted segments were inserted into pBluescript vector,and analyzed by restriction endonuclease mapping and DNA sequencing. RESULTS:Two clones were obtained. The sequence of one is coincidence with full mRNA sequence of mouse tuftelin,but the other lack the segment from 84 to 158.CONCLUSION: There are different mRNA transcripts of mouse tuftelin.