小鼠Dlx-5cDNA的克隆及序列测定

Cloning and sequencing of the coding region of mouse Dlx-5 gene

目的 :扩增小鼠Dlx 5的特异性片段 ,构建重组质粒并进行序列测定 ,为进一步的蛋白表达、抗体制备、基因转染等研究奠定基础。方法 :提取E14d小鼠的下颌弓组织总RNA ,以反转录cDNA为模板进行PCR反应 ,克隆Dlx 5至PMD 18T载体上并测序。结果 :从胎鼠下颌弓组织中克隆到Dlx 5基因片段约 869bp ,重组质粒经酶切显示获得正确重组子 ,测序结果与已知序列吻合。结论 :成功地从小鼠下颌弓中克隆到了Dlx 5基因 ,提示胎鼠的下颌弓组织中有Dlx 5基因的表达。

Objective:To clone the complete encoded sequence mouse Dlx-5 gene from mouse tooth germ, sequence and analyze its sequence.Method: Total RNA was isolated from embryo mouse mandible arch of 14 days and reverse-transcribed into cDNA.The desired cDNA was obtained by PCR. The segments was inserted into PMD-18 T vector and the positive clone was sequenced. Result: Dlx-5 cDNA was obtained from embryo mouse mandible arch. The sequence was consistent with that reported. One mutation base was found in this sequence.Conclusion: The Dlx-5 gene was sucessfully amplfied and sequenced. And this study provides the evidence of expression of Dlx-5 in embryo mouse mandible tissue.