摘要
目的检测Miyoshi肌病家系成员的DYSF基因突变,探讨变性高效液相色谱分析法(DHPLC)在基因研究中的效能。方法用RT-PCR技术扩增DYSF基因的编码序列,利用变性高效液相色谱分析技术(DHPLC)对PCR产物进行突变筛选,出现异常峰型的片段进行核苷酸序列测定,明确突变位点。结果患者DYSF基因存在6429delG突变。结论DHPLC能有效地用于基因突变的检测,该方法快速高效,可作为大样本筛查突变位点的一种便捷可靠手段。
Objective To identify the point mutation of DYSF gene in a miyoshi myopathy family and observe the value of denaturing high performance liquid chromatography (DHPLC). Methods mRNA from peripheral blood was amplified by the RT-PCR in a Chinese Miyoshi myopathy pedigree. The PCR products were analyzed by denaturing high-performance liquid chromatography (DHPLC). The samples with abnormal profiles were sequenced. Results Sequencing revealed a novel homozygous mutation, a 6 429delG, on exon 53 of the DYSF gene for the patients. Conclusion The DHPLC technique provides an efficient, sensitive and rapid method for detecting mutations of DYSFgene.
出处
《实用临床医药杂志》
CAS
2007年第4期43-45,共3页
Journal of Clinical Medicine in Practice