摘要
目的:了解OTOF基因在一常染色体显性遗传性听神经病家系的突变情况。方法:选择一个常染色体显性遗传听神经病家系中现存9名成员、3例散发听神经病患者和3名听力正常者为研究对象,用基因组DNA抽提试剂盒提取外周血DNA。对1例家系患者DNA进行OTOF基因全部编码区的PCR扩增,扩增产物经纯化后直接测序,测序结果与标准序列对照进行突变筛查;针对发现有突变的外显子,对其余受试者DNA样本进行PCR扩增和序列分析。结果:所有研究对象在OTOF基因相同部位上检测到10个新的碱基变异,但均未引起所编码氨基酸的改变。结论:该家系成员OTOF基因未发现有意义的突变位点,提示新基因参与家系耳聋的发生。
Objective: To investigate if the OTOF gene contributes to the non-syndromic hearing loss of a Chinese pedigree with dominantly inherited auditory neuropathy (AN). Method: The subjects included were 9 live individuals in an autosomal dominant AN pedigree, 3 sporadic AN patients and 3 normal-hearing controls. Genomic DNA was isolated from the peripheral leukocytes of the subjects using the Puregene DNA Isolation Kits. Firstly, the whole coding sequence of OTOF gene of one family patient were PCR amplified using specific primers. Each fragment was purified and subsequently analyzed by direct sequencing in an Applied Biosystems 3 730 automated DNA sequencer. The resultant sequence data were compared with the standard sequence to identify deafness-associated mutations. Other DNA samples were then screened for these mutations by PCR amplification and sequence analysis. Result:PCR amplifications were successfully conducted in all the subjects. Comparison of the resultant OTOF sequence in one family patient with the standard sequence identified 10 nucleotide variants which do not lead to amino acid change. These mutations were also detectable in other family individuals, 3 sporadic AN patients and 3 normal-hearing controls. Conclusion:The OTOF does not seem to contribute to the pathogenesis of this Chinese AN family, which suggest new gene(s) involvement.
出处
《临床耳鼻咽喉头颈外科杂志》
CAS
CSCD
北大核心
2007年第16期735-737,共3页
Journal of Clinical Otorhinolaryngology Head And Neck Surgery
基金
江苏省卫生厅医学科技发展基金(K200502)
江苏省自然科学基金(BK2005144)
江苏省"135工程"医学重点学科研究基金