本文選題：單核苷酸多態(tài)性 + 遺傳檢測��； 參考：《中國藥品生物制品檢定所》2006年碩士論文

【摘要】：遺傳檢測技術(shù)是遺傳質(zhì)量控制的重要手段之一，現(xiàn)有的檢測方法不能滿足科學(xué)發(fā)展的需要。近年來，隨著人類基因組研究的不斷深入，有關(guān)SNP的研究越來越受到重視，但小鼠SNP研究，特別是將SNP應(yīng)用于近交系小鼠遺傳檢測的研究比較少。為此，我們建立小鼠SNP遺傳檢測方法，并對SNP與生化標記位點多態(tài)性的關(guān)聯(lián)性進行研究，進一步完善我國實驗動物遺傳檢測技術(shù)，提高檢測水平。 本研究分為兩個部分：(1)小鼠SNP遺傳檢測方法的建立 采用單管雙向等位基因?qū)Ｒ恍詳U增(single-tube bi-directional allele specific amplification，SB-ASA)方法，，首先對5個國外品系的近交系小鼠的16個SNP位點進行了檢測，并進行測序驗證，同時設(shè)計雙盲實驗驗證該方法的可靠性。結(jié)果5個小鼠品系的16個SNP位點都成功地進行了分型，與測序的結(jié)果完全一致。雙盲實驗通過3個SNP位點綜合分析，可以準確地鑒別這5個小鼠品系。此外，我們對國內(nèi)培育的4個品系的近交系小鼠也進行了檢測，最終確定了4個品系的16個SNP位點等位基因，而且還繪制了9個品系小鼠的16個SNP位點的等位基因圖譜。這些研究不僅為各小鼠品系的遺傳鑒定和品系間的鑒別提供了實驗依據(jù)，而且還為近交系小鼠遺傳檢測提供了新方法。 (2)生化標記位點的分子生物學(xué)研究 本實驗嘗試從分子生物學(xué)水平去探索近交系小鼠生化標記位點多態(tài)性的機理，以及與SNP的關(guān)聯(lián)性。我們選擇了Car2、Gpi1和Hbb3個生化遺傳檢測位點，分別從生化電泳、DNA、RNA和翻譯蛋白4個方面進行了分析研究。結(jié)果發(fā)現(xiàn)Car2基因第38位Gin／His之間的轉(zhuǎn)換是引起所有小鼠品系形成Car2~a和Car2~b兩種電泳表型的原因；Gpi1基因的第247位氨基酸殘基Phe／Leu之間的轉(zhuǎn)換可能是形成Gpi1~a和Gpi1~b不同等位基因的原因；Hbb基因的第13、20和139位氨基酸殘基Cys／Gly、Ser／Ala和Thr／Ala之間的轉(zhuǎn)換是形成Hbb~d和Hbb~s兩種電泳表型的原因。 通過以上研究，建立了近交系小鼠的SNP遺傳質(zhì)量檢測方法，確定了國內(nèi)4個品系的16個SNP位點等位基因，繪制了9個品系小鼠的SNP位點等位基因圖譜。同時，通過分子生物學(xué)手段對生化標記位點多態(tài)性的形成機制進行研究，初步闡明三個生化位點多態(tài)的形成機理。本研究對提高我國實驗動物遺傳檢測技術(shù)水平，促進生命科學(xué)發(fā)展具有重要意義。
[Abstract]:Genetic detection technology is one of the important means of genetic quality control, the existing detection methods can not meet the needs of scientific development. In recent years, with the development of human genome research, more and more attention has been paid to the study of SNP, but the study of mouse SNP, especially the application of SNP to the genetic detection of inbred mice, is less. Therefore, we established the method of SNP genetic detection in mice, and studied the association between SNP and polymorphism of biochemical marker loci in order to further improve the genetic detection technology of laboratory animals in China and improve the detection level. This study was divided into two parts: 1) Establishment of SNP genetic detection method in mice using single-tube bi-directional allele specific amplification SB-ASAA method. First, 16 SNP loci of 5 inbred strains of foreign strains were detected. The reliability of the method was verified by sequencing and double-blind experiments were designed to verify the reliability of the method. Results 16 SNP loci of 5 mouse strains were successfully typed, and the results were in good agreement with the sequencing results. Three SNP loci were used to identify the five mouse strains. In addition, 16 SNP loci alleles of 4 strains were determined and alleles of 16 SNP loci of 9 strains were plotted. These studies not only provide experimental basis for genetic identification and identification among strains of mice, but also provide a new method for genetic detection of inbred mice. (2) Molecular biology of biochemical marker loci in this experiment, we try to explore the mechanism of the polymorphism of biochemical marker loci in inbred mice and its association with SNP from the molecular biological level. We selected Car2Gpi1 and Hbb3 sites for biochemical genetic analysis, and analyzed them from four aspects of biochemical electrophoretic DNA RNA and translation protein, respectively. It was found that the conversion between the 38th Gin/His of the Car2 gene was the cause of the formation of two electrophoretic phenotypes of Car2~a and Car2~b in all mouse strains. The conversion between the 247th amino acid residues Phe/Leu of the Gpi1 gene may be the formation of different alleles of Gpi1~a and Gpi1~b. The conversion between Cys / Glycine Serr / Ala and Thr/Ala in the 1320 and 139 amino acid residues of HB gene was responsible for the formation of two electrophoretic phenotypes of Hbb~d and Hbb~s. Based on the above studies, the SNP genetic quality detection method of inbred strain mice was established, 16 alleles of SNP loci in 4 strains in China were determined, and the allelic map of SNP loci in 9 strains of mice were plotted. At the same time, the formation mechanism of the polymorphism of biochemical marker loci was studied by molecular biology, and the formation mechanism of the polymorphism of three biochemical loci was preliminarily elucidated. This study is of great significance to improve the technology of genetic detection of laboratory animals and promote the development of life science in China.
【學(xué)位授予單位】：中國藥品生物制品檢定所
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2006
【分類號】：R-332

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