SD大鼠Atoh1基因CDS区的克隆及序列分析  

The cloning and sequencing of SD rat Atoh1 gene CDS region

作　　者：郑国玺[1] 祝康[1] 朱珠[1] 侯瑾[1] 韦俊荣[1] 许珉[1] 

机构地区：[1]西安交通大学医学院第二附属医院耳鼻喉病院,陕西西安710004

出　　处：《西安交通大学学报（医学版）》2011年第4期466-469,共4页Journal of Xi’an Jiaotong University（Medical Sciences）

基　　金：国家自然科学基金资助项目(No.30471877);陕西省科技攻关项目(No.2010K15-08)~~

摘　　要：目的利用基因工程方法克隆SD大鼠Atoh1 cDNA编码序列并测定分析其克隆序列。方法采用非对称互补引物/模板法,制备Atoh1 cDNA的编码序列,将其克隆入PMD-19T载体中并测序。结果经酶切鉴定、测序分析表明,扩增得到的大鼠Atoh1基因CDS区全长为1 056 bp,编码351个氨基酸;测定序列与GenBank中公布的参考序列对比,有2处碱基发生突变,但克隆序列编码的氨基酸序列与参考序列完全一致。这两处碱基应为单核苷酸多态性(SNP)位点,突变为无义突变,不影响蛋白的表达。结论成功克隆了Atoh1 cDNA编码序列,为进一步对感音神经性耳聋的基因治疗奠定了基础。Objective To make use of gene engineering technique to clone and analyze the SD rats Atoh1 cDNA coding sequence.Methods By means of asymmetrical primer/ template,double stranded cDNA of Atoh1 was obtained.Then the cDNA coding sequence was subcloned into PMD-19T vector and sequenced.Results Evidence of DNA sequence analysis and restriction enzyme digestion showed that the rat Atoh1 gene amplified length of CDS area was 1056bp and 351 amino acids were encoded.By comparing it with the reference sequences published in GeneBank,we found two base mutations in the sequence;it may be single nucleotide polymorphisms in the nucleotide to induce nonsense mutation;thus we can deduce that amino acid of cloning sequences is the same as that of the reference sequences.Conclusion Atoh1 cDNA coding sequence was cloned successfully,which will guide further research on gene therapy for sensorineural hearing loss.

关 键 词：感音神经性耳聋 Atoh1 基因克隆 耳蜗毛细胞 

分 类 号：R764.4[医药卫生—耳鼻咽喉科]