CAJ | 학술논문

目的探讨COL1A1基因中一个新突变c.3208G＞A导致Ⅰ型成骨不全的致病机理.方法应用标准的方法提取外周血全基因组DNA,PCR扩增基因的整个编码区域和内含子-外显子边界,应用ABI 3100/3130测序仪对PCR产物进行直接测序.从患者的EB病毒转化永生B细胞系中提取全RNA,用寡(dT) 18引物合成第一链cDNA,对PCR产物直接测序或者用TA克隆质粒DNA测序.结果在一个Ⅰ型成骨不全大家庭中检测到一种位于COL1A1基因上的新突变c.3208G＞A,为剪接突变.结论发现了COL1A1基因中的新的剪接突变c.3208G＞A.
목적 탐토COL1A1기인중일개신돌변c.3208G＞A도치Ⅰ형성골불전적치병궤리.방법 응용표준적방법제취외주혈전기인조DNA,PCR확증기인적정개편마구역화내함자-외현자변계,응용ABI 3100/3130측서의대PCR산물진행직접측서.종환자적EB병독전화영생B세포계중제취전RNA,용과(dT) 18인물합성제일련cDNA,대PCR산물직접측서혹자용TA극륭질립DNA측서.결과 재일개Ⅰ형성골불전대가정중검측도일충위우COL1A1기인상적신돌변c.3208G＞A,위전접돌변.결론 발현료COL1A1기인중적신적전접돌변c.3208G＞A.
Objective To investigate the genetic cause for a large family affected with type Ⅰ osteogenesis imperfecta.Methods Genomic DNA was extracted from peripheral venous blood samples.The entire coding region and intron-exon boundaries of the COL1A1 gene were subjected to PCR amplification and direct sequencing.Total RNA was also extracted from immortalized B cell lines from the patients,with the first strand of cDNA synthesized with an oligo(dT)18 primer.The PCR products were directly sequenced using the TA cloned plasmid.Results A c.3208G＞ A mutation has been identified in the COL1A1 gene,which can alter the splicing pattern of mRNA.Conclusion A novel splicing mutation c.3208G＞A of the COL1A1 gene probably underlies the disease.