CAJ | 학술논문

目的对1个有3例女性杂合子患者的X-连锁肾上腺-脑白质营养不良(X-linked adrenoleukodystrophy,X-ALD)家系进行基因突变分析.方法提取1例患者的外周血白细胞总RNA,以此为模板,采用长链逆转录-PCR技术,分4个片段扩增ABCD1基因(X-ALD疾病基因)编码序列并对其PCR产物进行直接测序.通过PCR和限制性内切酶分析患者及其家庭成员相应的基因组DNA片段,进一步确证所发现的基因突变.对突变及其对ALD蛋白结构的影响进行生物信息学分析.结果在患者A月CD1基因第1外显子的第283位密码子发现1个新的错义突变:CAC→CGC(p.H283R),此突变使相应DNA片段的1个Msl Ⅰ酶切位点消失,其子不存在此突变.突变所改变的氨基酸残基在进化上高度保守.p.H283R突变位于ALD蛋白的跨膜结构域,可引起该分子跨膜区整体结构的改变.结论在国内首次报道了3例杂合子女性X-ALD患者,并在其家系发现了1个新的ABCD1基因突变,即p.H283R突变.
목적 대1개유3례녀성잡합자환자적X-련쇄신상선-뇌백질영양불량(X-linked adrenoleukodystrophy,X-ALD)가계진행기인돌변분석.방법 제취1례환자적외주혈백세포총RNA,이차위모판,채용장련역전록-PCR기술,분4개편단확증ABCD1기인(X-ALD질병기인)편마서렬병대기PCR산물진행직접측서.통과PCR화한제성내절매분석환자급기가정성원상응적기인조DNA편단,진일보학증소발현적기인돌변.대돌변급기대ALD단백결구적영향진행생물신식학분석.결과 재환자A월CD1기인제1외현자적제283위밀마자발현1개신적착의돌변:CAC→CGC(p.H283R),차돌변사상응DNA편단적1개Msl Ⅰ매절위점소실,기자불존재차돌변.돌변소개변적안기산잔기재진화상고도보수.p.H283R돌변위우ALD단백적과막결구역,가인기해분자과막구정체결구적개변.결론 재국내수차보도료3례잡합자녀성X-ALD환자,병재기가계발현료1개신적ABCD1기인돌변,즉p.H283R돌변.
Objective To identify ABCD1 gene mutation in a Chinese family with three heterozygous female patients. Methods Four fragments covering the entire coding sequence of the ABCD1 gene from one of the female patients were amplified by reverse transcription-PCR. The PCR products were directly sequenced. The result of sequencing was confirmed by restriction enzyme digestion of PCR products from genomic DNA. Human ABCD1 gene and ALD protein were aligned with those of rat, monkey, mouse and cattle by Clustal X 1.83. Softwares of Motif Scan, TMpred and ESYpred3D were used to predict the effect of the mutation on the structure of the ALD protein. Results A novel missense mutation, CAC to CGC,was found at codon 283 of the ABCD1 gene from the patient, resulting in the replacement of histidine by arginine. This mutation abolished an Msl Ⅰ site in the gene. Her son was free from this mutation. The mutated amino acid residue (283H) was highly conservative in evolution, and the mutation caused a dramatic change in the structure of the ALD protein. Conclusion Three female patients heterozygous for ABCD1 gene mutation were first reported in China, and a novel mutation, p. H283R, was identified in this X-ALD family.