【摘要】： 目的研究D2S1399和D5S2500基因座在中國(guó)華東地區(qū)漢族群體的遺傳多態(tài)性,評(píng)估其在法科學(xué)中的應(yīng)用價(jià)值;利用重組質(zhì)粒制備這兩個(gè)STR基因座的分型標(biāo)準(zhǔn)物。 方法應(yīng)用PCR、聚丙烯酰胺垂直電泳及銀染技術(shù)檢測(cè)D2S1399和D5S2500基因座的遺傳多態(tài)性。從聚丙烯酰胺凝膠中分離出經(jīng)PCR擴(kuò)增后的等位基因片段,二次擴(kuò)增并純化后的等位基因片段被分別插入到PUC質(zhì)粒載體中。利用DNA測(cè)序證實(shí)插入片段大小及結(jié)構(gòu)的正確性后,用國(guó)際標(biāo)準(zhǔn)將插入的等位基因片段進(jìn)行命名。再轉(zhuǎn)染到適當(dāng)?shù)腅. coli DH5αTM細(xì)胞內(nèi)選擇培養(yǎng),以純化質(zhì)粒為模板,擴(kuò)增出各基因座的等位基因片段后混合獲得等位基因分型標(biāo)準(zhǔn)物。 結(jié)果D2S1399和D5S2500基因座在中國(guó)華東漢族群體分別檢出11個(gè)和9個(gè)等位基因,其觀察雜合度分別為0.745和0.807,多態(tài)信息含量(PIC)分別為0.850和0.750,個(gè)體識(shí)別率分別為0.958和0.917,非父排除率(PE)分別為0.554和0.643。成功地建立了能大量生產(chǎn)的D2S1399及D5S2500基因座的標(biāo)準(zhǔn)等位基因片段的重組質(zhì)粒庫(kù),并制備出大量?jī)?yōu)質(zhì)分型標(biāo)準(zhǔn)物。 結(jié)論D2S1399和D5S2500基因座是高度多態(tài)性的STR基因座,在法醫(yī)學(xué)個(gè)人識(shí)別和親子鑒定中有重要應(yīng)用價(jià)值。通過(guò)分子克隆制備D2S1399及D5S2500基因座的等位基因分型標(biāo)準(zhǔn)物是一種實(shí)現(xiàn)D2S1399及D5S2500基因座分型標(biāo)準(zhǔn)化的可行方法。
[Abstract]:Objective to study the genetic polymorphisms of D2S1399 and D5S2500 loci in the Han population of East China, to evaluate their application value in French science, and to prepare the standard compounds of the two STR loci by using recombinant plasmids. Methods PCR, polyacrylamide vertical electrophoresis and silver staining were used to detect the genetic polymorphism of D2S1399 and D5S2500 loci. The alleles amplified by PCR were isolated from the polyacrylamide gel and inserted into the PUC plasmid vector after secondary amplification and purification. After DNA sequencing was used to confirm the correctness of the size and structure of the inserted fragment, the inserted allele fragment was named by international standard. The alleles of each locus were amplified by using purified plasmids as template, and then the alleles were mixed to obtain the standard alleles. Results 11 and 9 alleles of D2S1399 and D5S2500 loci were detected in the Han population of East China respectively. The observed heterozygosity was 0.745 and 0.807, and the polymorphic information content (PIC) was 0.850 and 0.750, respectively. The individual recognition rates were 0.958 and 0.917, and the non-paternal exclusion rates (PE) were 0.554 and 0.643, respectively. A recombinant plasmid library of standard allelic fragments of D2S1399 and D5S2500 loci was successfully constructed and a large number of high quality genotyping standard materials were prepared. Conclusion D2S1399 and D5S2500 loci are highly polymorphic STR loci, which have important application value in forensic personal identification and paternity identification. The preparation of allelic alleles of D2S1399 and D5S2500 loci by molecular cloning is a feasible method for the standardization of D2S1399 and D5S2500 loci.
【學(xué)位授予單位】：蘇州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2007
【分類號(hào)】：R394

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1 夏水秀;華東地區(qū)漢族人群D2S1399和D5S2500基因座的遺傳多態(tài)性研究及其等位基因分型標(biāo)準(zhǔn)物的構(gòu)建[D];蘇州大學(xué);2007年

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