小鼠成釉蛋白基因cDNA全序列的克隆

Cloning of full length cDNA sequence of the mouse ameloblastin

目的 筛选与小鼠牙胚发育相关的特异基因。方法 利用随机引物、反转录酶合成特异和非特异探针 ,采用差异显示方法筛选小鼠牙胚cDNA文库 ,挑选阳性克隆并测序。结果 得到 6个阳性克隆 ,经测序证实其中 1个克隆序列与大鼠成釉蛋白序列高度同源 :其中用pTriplEX 3′引物测得的 5 2 6个碱基序列与大鼠成釉蛋白基因 5′端的 32 5 80序列有 4 97个碱基一致 ,5′引物测得的 5 6 7个碱基序列与大鼠成釉蛋白基因 3′端的 12 85 185 4序列有 5 33个碱基一致。结论 筛选到小鼠釉基质特异蛋白———成釉蛋白基因的cDNA全序列。

Objective Screening for special genes of matrix proteins of dentin and enamel of mouse dental germ. Methods A cDNA library of dental germ of mouse was screened by differential display. The interesting clones were sequenced. Results Six positive clones were isolated from the cDNA library. The sequence of one of the six positive clones was homologous with the ameloblastin sequence of rat. There are 497 homologous base pairs between the 526 base pairs sequenced by pTriplEX 3′ primer of this clone and the 32 580 sequence of the rat ameloblastin gene; and there are 533 homologous base pairs between the 567 base pairs sequenced by pTriplEX 5′ primer of this clone and the 1285 1854 sequence of the rat ameloblastin gene. Conclusions The full length cDNA sequence of the mouse ameloblastin was cloned.