本文選題：家族性腺瘤息肉病 + APC��； 參考：《南京醫(yī)科大學(xué)》2015年碩士論文

【摘要】：目的:通過(guò)基因診斷技術(shù)尋找和分析家族性腺瘤息肉病(familial adenomatous polyposis,FAP)家系結(jié)腸腺瘤息肉病(adenomatous polyposis coli,APC)基因致病性胚系突變及其特點(diǎn),同時(shí)檢測(cè)基于多重連接依賴(lài)性探針擴(kuò)增(multiplex ligationdependent probe amplification,MLPA)和變性高效液相色譜技術(shù)(denaturing High Performance Liquid Chromatography,DHPLC)技術(shù)平臺(tái)的基因診斷效力。方法:提取2005年至2013就診于我院的14個(gè)FAP家系成員外周血DNA,聯(lián)合應(yīng)用MLPA、PCR-DHPLC技術(shù)及直接測(cè)序等方法對(duì)APC基因胚系突變進(jìn)行檢測(cè),尋找各家系A(chǔ)PC基因致病性胚系突變。隨后分析突變位點(diǎn)周?chē)鷫A基序列,探索APC基因胚系突變特點(diǎn)。結(jié)果:14個(gè)家系中有6個(gè)家系篩查出了APC基因胚系突變,分別是c.5432CT(p.Ser1811Leu)、兩個(gè)c.3926_3930del AAAAG(p.Glu1309Aspfs X4)、c.3921_3924del AAAA(p.Ile1307Metfs X13)、c.3184_3187del CAAA(p.Gln1061Aspfs X59)和c.4127_4126del AT(p.Tyr1376Lysfs X9),所有的缺失突變都導(dǎo)致了截?cái)嗟鞍椎漠a(chǎn)生。14個(gè)家系中均未發(fā)現(xiàn)有大片段缺失與重復(fù)。分析缺失突變位點(diǎn)周?chē)鷫A基序列發(fā)現(xiàn),c.3926_3930del AAAAG與c.3921_3924del AAAA位于A(yíng)AAAG短串聯(lián)重復(fù)序列中,c.3184_3187del CAAA位于間斷的短串聯(lián)重復(fù)序列中,而c.4127_4128del AT則位于5'-CCTGAACA-3',3'-ACAAGTCC-5回文序列(反向重復(fù)序列),并且大部分的突變位于密碼子1309周?chē)�。結(jié)論:包含短串聯(lián)重復(fù)序列及回文序列的區(qū)域?yàn)锳PC基因致病性胚系突變高發(fā)區(qū)域,以小片段缺失為主,尤其是密碼子1309位點(diǎn)的5堿基缺失最為常見(jiàn)�；贛LPA和DHPLC技術(shù)平臺(tái)的基因診斷技術(shù)可快速、高效地進(jìn)行基因突變篩查。
[Abstract]:Objective: to identify and analyze the mutation and characteristics of pathogenic embryoid line of adenomatous polyposis in a family of familial adenomatous polyposis (adenomatous polyposis) by means of gene diagnosis technique. At the same time, the gene diagnostic efficacy of multiplex ligationdependent probe amplification (MLPA) and denaturing High Performance Liquid chromatography (DHPLC) platform based on multiplex ligationdependent probe amplification and denaturing high performance liquid chromatography (denaturing High Performance Liquid) were also detected. Methods: peripheral blood DNA was extracted from 14 FAP family members from 2005 to 2013 in our hospital. APC gene embryogenic mutation was detected by MLPA-PCR-DHPLC and direct sequencing. The pathogenicity of APC gene was found in all families. Then the base sequence around the mutation site was analyzed to explore the characteristics of APC gene blastocyst mutation. Results: in 6 of 14 families, APC gene embryonal mutations were screened. The results were as follows: C. 5432CTp.Ser1811 Leuer, two c.3926_3930del AAAAG(p.Glu1309Aspfs X4s, c. 3921 3924del AAAA(p.Ile1307Metfs X13 / c. 31844 del CAAA(p.Gln1061Aspfs X59) and c.4127_4126del AT(p.Tyr1376Lysfs X9, all deletion mutations resulted in the production of truncated proteins. No large deletion or duplication was found in 14 families. By analyzing the base sequence around the deletion mutation site, we found that AAAAG and c.3921_3924del AAAA are located in the AAAAG short tandem repeat sequence, and the c. 3184 CAAA is located in the discontinuous short tandem repeat sequence. The c.4127_4128del AT was located in the palindrome sequence (reverse repeat sequence) of 5CT-CCGAACA-3ACAAGTCC-5, and most of the mutations were located around codon 1309. Conclusion: the region containing short tandem repeats and palindromes is a high incidence region of APC gene pathogenicity embryo line mutation. Small fragment deletion is the most common, especially the 5-base deletion at codon 1309. Gene diagnosis technology based on MLPA and DHPLC technology platform can quickly and efficiently screen gene mutation.
【學(xué)位授予單位】：南京醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類(lèi)號(hào)】：R735

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 劉曉蓉,單祥年,FriedlW,UhlhaasS,李金田,ProppingP,王亞平;應(yīng)用蛋白截短技術(shù)檢測(cè)APC基因胚系突變[J];遺傳學(xué)報(bào);2005年09期

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本文編號(hào)：1825845