中华医学遗传学杂志
中華醫學遺傳學雜誌
중화의학유전학잡지
CHINESE JOURNAL OF MEDICAL GENETICS
2008年
2期
164-167
,共4页
家族性腺瘤性息肉病%变性高效液相色谱%结肠腺瘤性息肉病基因%突变
傢族性腺瘤性息肉病%變性高效液相色譜%結腸腺瘤性息肉病基因%突變
가족성선류성식육병%변성고효액상색보%결장선류성식육병기인%돌변
familial adenomatous polyposis%denaturing high performance liquid chromatography%adenomatous polyposis coli gene%mutatio
目的 应用变性高效液相色谱(denaturing high performance liquid chromatography,DHPLC)技术检测我国家族性腺瘤性息肉病(familial adenomatons polyposis,FAP)家系的结肠腺瘤性息肉病(adenoinatous pelyposis coli,APC)基因变异特征,研究其病因机制.方法 采集31个家系的先证者、患者和家系成员的外周血淋巴细胞,抽提DNA并以降落式PCR扩增APC基因各外显子和启动子.基因突变检测先由DHPLC进行筛选,发现异常峰者进行测序鉴定并TA克隆鉴定,结果与网络数据进行比对.结果 31个家系中共有15个家系检出了12种不同的突变类型,FAP家系APC基因的突变检出率为48.39%.发现了4种新的突变及3例不同的内含子突变.4个新的突变分别位于255、677、1192、1403密码子,均为移码突变.证明了DHPLC能检出APC基因的突变.在APC基因的突变中,移码突变占86.67%,无义突变占13.33%,说明移码突变是中国人APC基因突变的主要方式.在突变位点上,第15外显子突变最常见,约占86.67%.结论 FAP家系APC基因的突变检出率为48.39%,发现了4种新的导致蛋白编码改变的突变.证实中国人FAP家系中APC基因突变位点以第15外显子最常见,类型以移码突变为主.
目的 應用變性高效液相色譜(denaturing high performance liquid chromatography,DHPLC)技術檢測我國傢族性腺瘤性息肉病(familial adenomatons polyposis,FAP)傢繫的結腸腺瘤性息肉病(adenoinatous pelyposis coli,APC)基因變異特徵,研究其病因機製.方法 採集31箇傢繫的先證者、患者和傢繫成員的外週血淋巴細胞,抽提DNA併以降落式PCR擴增APC基因各外顯子和啟動子.基因突變檢測先由DHPLC進行篩選,髮現異常峰者進行測序鑒定併TA剋隆鑒定,結果與網絡數據進行比對.結果 31箇傢繫中共有15箇傢繫檢齣瞭12種不同的突變類型,FAP傢繫APC基因的突變檢齣率為48.39%.髮現瞭4種新的突變及3例不同的內含子突變.4箇新的突變分彆位于255、677、1192、1403密碼子,均為移碼突變.證明瞭DHPLC能檢齣APC基因的突變.在APC基因的突變中,移碼突變佔86.67%,無義突變佔13.33%,說明移碼突變是中國人APC基因突變的主要方式.在突變位點上,第15外顯子突變最常見,約佔86.67%.結論 FAP傢繫APC基因的突變檢齣率為48.39%,髮現瞭4種新的導緻蛋白編碼改變的突變.證實中國人FAP傢繫中APC基因突變位點以第15外顯子最常見,類型以移碼突變為主.
목적 응용변성고효액상색보(denaturing high performance liquid chromatography,DHPLC)기술검측아국가족성선류성식육병(familial adenomatons polyposis,FAP)가계적결장선류성식육병(adenoinatous pelyposis coli,APC)기인변이특정,연구기병인궤제.방법 채집31개가계적선증자、환자화가계성원적외주혈림파세포,추제DNA병이강락식PCR확증APC기인각외현자화계동자.기인돌변검측선유DHPLC진행사선,발현이상봉자진행측서감정병TA극륭감정,결과여망락수거진행비대.결과 31개가계중공유15개가계검출료12충불동적돌변류형,FAP가계APC기인적돌변검출솔위48.39%.발현료4충신적돌변급3례불동적내함자돌변.4개신적돌변분별위우255、677、1192、1403밀마자,균위이마돌변.증명료DHPLC능검출APC기인적돌변.재APC기인적돌변중,이마돌변점86.67%,무의돌변점13.33%,설명이마돌변시중국인APC기인돌변적주요방식.재돌변위점상,제15외현자돌변최상견,약점86.67%.결론 FAP가계APC기인적돌변검출솔위48.39%,발현료4충신적도치단백편마개변적돌변.증실중국인FAP가계중APC기인돌변위점이제15외현자최상견,류형이이마돌변위주.
Objective To analyze the adenomatous polyposis coli (APC)gene mutations in familial adenomatous polyposis(FAP)in Chinese.Methods DNA was extracted from blood samples taken from 31 FAP families ,and all formance liquid chromatography followed by sequencing if abnormal profile Was detected.Results Twelve categories ofAPC gene mutations were found in 15 FAP families(48.39%)including 4 novel mutations in coding region and 3 mutations in introns.The 4 novel mutations in coding region were frameshift mutations and located in codons 255,677,1192 and 1403 respectively.Most mutations were clustered in exon 15 of APC gene leading to frameshift and accounted for 86.67%.Others were nonsense mutatiom(13.33%).Conclusion The mutation rate of the APC gene in this group of Chinese FAP families was about 48.39%,and 4 novel mutations were detected.Frameshift mutation was the major mutation type in Chinese FAr and mainly located in exon 15.
