中国人Wilson病患者ATP7B基因外显子突变研究

Mutations in exon of ATP7B gene in Chinese patients with Wilson's disease

目的对Wilson病(WD)患者ATP7B基因外显子进行PCR扩增测序,研究其突变的特点。方法对41例患者(WD组)、10例健康者(对照组)以及1个WD家系(先证者女儿及其父母3人)提取基因组DNA,PCR扩增外显子相关片段,并对扩增产物进行直接测序。结果健康对照组未见异常,WD组发现11例患者存在外显子8点突变,其中6例患者呈Arg778Leu的复合杂合突变;4例患者存在外显子12点突变,其中2例存在Arg952Lys突变。在Wilson病家系中,先证者女儿携带2种杂合性突变,分别是父源的外显子8中Arg778Leu杂合性突变和母源的外显子13中Pro992Leu杂合性突变,其父母均为表型正常的杂合子携带者。结论中国人WD患者中ATP7B基因外显子8、12为突变的热点区,但也存在其他外显子的突变,如外显子13。

Objective To amplify the ATP7B gene of Wilson disease（WD） patients by PCR and to sequence the amplification product,so as to characterize the possible mutations.Methods The genomic DNA of 41 WD patients,10 normal controls and a WD genealogy（proband＇s daughter and parents） were extracted.The fragments of exon 8 and 12 of ATP7B gene were amplified using PCR,and the PCR products were directly sequenced.Results No abnormality was found in the control group.Mutations of exon 8 were found in 11 WD patients,with 6 WD patients having Arg778Leu heterozygous mutations.Mutations of exon 12 were found in 4 WD patients,with 2 patients having Arg952Lys mutations.In the sibs of the WD patient,the proband＇s daughter carried 2 heterozygous mutations：Arg778Leu mutation of exon 8 was from her father and Pro992Leu mutation of exon 13 from her mother;the proband＇s parents were found as normal heterozygous carriers.Conclusion Exon 8 and 12 of ATP7B gene are prone to mutations in Chinese WD population,and the mutations of other exons,such as exon 13,also exist.