中国瑶族人群(广西)9个STR基因多态性研究

采用STR基因扫描技术选择D3S1358,vWA,FGA,THO1,TPOX,CSF1P0,D5S818, D13S317 和 D7S820 9种STR基因座,研究我国瑶族人群STR遗传多态性。在瑶族群体中9个STR基因座共检出61个等位基因,其频率分布在0.0054~0.5924,平均杂合度为0.7357,多态信息量为0.6887,累积个体识别率为2.02×10-10,非父排除率为0.9999。结果表明,在人类遗传学、法学科等领域建立本民族本地区遗传学资料是十分重要和必不可少的。 Study on 9 STR Loci Polymorphism from Chinese Yao Ethnic Group(Guangxi) GAO Fang1,BI Shi-hua2,LAI Jiang-hua1,LI Sheng-bin1,2 1.National Laboratory of Forensic Sciences,Xian Jiaotong University,Xi'an 710061; 2.Human Genome Genter,Institute of Genetics and Developmental Biology,Chinese Academy of Sciences,Beijing 100101 China Abstract:Genetic polymorphism of nine STR loci was investigated from a Chinese Yao population based on STR Genescan.Sixty one alleles was determined for 9 loci,such as D3S1358,vWA,FGA,THO1,TPOX,CSF1P0,D5S818,D13S317 and D7S820 with their frequencies 0.0054~0.5924.The average heterozygosity(H) was 0.7357,polymorphism information content(PIC) was 0.6887,the accumulative discrimination power(DP) was 2.02×10-10 and the probability of paternity exclusion(PPE) was 0.9999.These results suggested that the nine STR loci are very useful for human identification,such as analyzing forensic casework,establishing DNA databases,processing paternity test and studying gene natural resources. Key words：STR;genescan;Yao ethnic group;individual identification     

用多重PCR检测上海地区汉族人群9个STR基因座的多态性

利用多重PCR和四色荧光（5-FAM,JOE,NED和ROX）自动化检测技术调查上海地区汉族人群D3S1358、vWA、FGA、D8S1179、 D21S11、 D18S51、D5S818、D13S317、D7S820等9个STR基因座多态性分布并计算该9个基因座的基因频率（Pi）、个体鉴别力（DP）、无偏倚期望杂合性（H）、多态性信息含量（PIC）和非父排除概率（PE）。结果显示：9个STR基因座的基因型分布符合Hardy-Weinberg平衡,9个STR基因座中FGA基因座的DP值最高为0.9584,D8S1179的H值最高为0.9403,D18S51的PIC值最高为0.8560,D18S51的PE值最高为0.7391,9个STR基因座累积个体鉴别力(CDP)为0.9999996,累积非父排除能力(CPE)为0.99991。9个STR基因座适合作为中国人群的遗传标志,用于人类学、遗传疾病基因连锁分析、法医学亲子鉴定和个体识别等研究领域。 Abstract:By multiplex amplification and four fluorescent technique,the polymorphism distributions of nine STR loci,D3S1358,vWA,FGA,D8S1179,D21S11,D18S51,D5S818,D13S317 and D7S820 were investigated in Shanghai Han population.Gene frequency (Pi),power of discrimination (DP),polymorphism information content (PIC) expected heterozygosity (H) and probability of paternity exclusion (PE) were calculated.All loci meet Hardy-Weinberg equilibrium.DP of FGA locus,H of D8S1179 locus,PIC of D18S51 locus and PE of D18S51 locus are the biggest among nine STR loci.Cumulate DP (CDP) of nine STR loci is 0.9999996,Cumulate PE (CPE) of nine STR loci is 0.99991.Nine STR loci could be used as the genetic markers of Chinese population in the studies of anthropology,linkage analysis of genetic disease genes,individual identification and paternity test in forensic medicine.     

贵州地区汉族人群THO1、TPOX、CSF1PO基因座的遗传多态性

为了解贵州地区汉族群体中THO1、TPOX、CSF1PO基因座的遗传多态性,获得这3个基因座的群体遗传学数据和法医学相关数据。采自贵州地区汉族无关个体的110份EDTA抗凝血样用Chelex法提取DNA,应用PCR复合扩增技术扩增样本后,聚丙烯酰胺凝胶电泳分型。对3个STR基因座的等位基因频率进行了调查分析,并与其他汉族人群的等位基因频率进行了比较。在贵州汉族群体中,3个基因座的基因型分布符合Hardy-Weinberg平衡。3个STR基因座总个体识别率为0.9986,累积非父排除率为0.832。表明这3个基因座在法医学个体识别及亲子鉴定中是很有价值的遗传标记系统。 Abstract:To understand the genetic polymorphism at THO1,TPOX,CSF1PO STR loci for Han population in Guizhou Province,and construct a preliminary database,EDTA-blood specimens were collected from the 110 unrelated individuals in Han population from Guizhou.The DNA samples were extracted with Chelex method and amplified by multiplex polymerase chain reaction.The PAGE was used to type the PCR products.The allele frequencies were compared with other Han populations.The genotype distributions of THO1,TPOX and CSF1PO were in accordance with Hardy-Weinberg equilibrium.The combined PD and PE were 0.9986 and 0.832 respectively.All of the three loci in this study provide useful marker for forensic paternity test and individual identification.     

Polymorphism profile of nine short tandem repeat Loci in the Han chinese

Nine short tandem repeat (STR) markers (D3S1358, VWA, FGA, THO1, TPOX, CSFIPO, D5S818, D13S317, and D7S820) and a sex-identification marker (Amel-ogenin locus) were amplified with multiplex PCR and were genotyped with a four-color fluorescence method in samples from 174 unrelated Han individuals in North China. The allele frequencies, genotype frequencies, heterozygosity, probability of discrimination powers, probability of paternity exclusion and Hardy-Weinberg equilibrium expectations were determined. The results demonstrated that the genotypes at all these STR loci in Han population conform to Hardy-Weinberg equilibrium expectations. The combined discrimination power (DP) was 1.05×10-10 within nine STR loci analyzed and the probability of paternity exclusion (EPP) was 0.9998. The results indicate that these nine STR loci and the Amelo-genin locus are useful markers for human identification, paternity and maternity testing and sex determination in forensic sciences.     

STR遗传多态性研究中样本数量对等位基因检出数量的影响

以30个不同民族9个常染色体STR基因座(D3S1358, vWA, FGA, TH01,TPOX, CSF1PO, D13S317, D5S818, D7S820)的群体遗传研究数据资料为例, 探讨群体遗传学研究中常染色体STR基因座等位基因检出数量与样本量之间的关系, 即样本量对等位基因检出数量的影响。结果显示, 在一定范围之内, 样本量的大小与所观测到的不同基因座等位基因检出数量之间存在正相关关系。当超过一定范围时, 样本量的继续增加不再明显影响等位基因的检出数量。杂合度较低的位点随样本量的变化波动较大, 杂合度较高的位点随样本量的变化波动较小。     

Genetic analysis of 15 STR loci in Chinese Han population from West China

Allele frequencies for 15 short tandem repeat （STR） loci （D8S1179, D21S11, D7S820, CSFIPO, D3S1358, TH01, D13S317, D16S539, D2S1338, D19S433, vWA, TPOX, D18S51, D5S818, and FGA） were obtained from 7,636 unrelated individuals of Chinese Han population living in Qinghai and Chongqing, China. Totally 206 alleles were observed, with the corresponding allele frequencies ranging from 0.0001-0.4982. Chi-square test showed that all of the STR loci agreed with the Hardy-Weinberg equilibrium. We also compared our data with previously published population data of other ethnics or areas. The results are valuable for human identification and paternity testing in Chinese Han population.     

中国朝鲜族9个STR基因座遗传多态性研究

为丰富中华民族基因数据库,获取中国吉林省特有少数民族--朝鲜族D3S1358、vWA、FGA、TH01、TPOX、CSF1PO、D5S818、D13S317、D7S820等9个STR基因座的群体遗传数据。采用四色荧光标记STR基因扫描技术,检测91个无关个体血液样本。结果共检出81种等位基因,其基因频率分布在0.0055~0.4615之间;共检出196种基因型,其基因型频率分布在0.0110~0.9890之间。9个STR基因座基因型频率观察值与期望值均符合Hardy-Weinberg平衡定律（P＞0.05）。9个基因座的多态信息量PIC（polymorphic information content）分布于0.6863~0.8807之间,杂合度H（heterozygosity）分布于0.6919~0.8809之间,个体识别力DP（discrimination power）分布于0.8301~0.9670之间,非父排除率PPE（probability of paternity exclusion）分布于0.8590~0.9942之间。研究结果可应用于人类群体遗传学及法医学研究等领域。 Genetic Polymorphism of 9 STR loci in Chaoxian National Minority of China GAO Ya1,JIN Tian-bo1,LAI Jiang-hua1,CHEN Teng1,ZHENG Hai-bo1,ZHU Bo-feng1,HU Song-nian2,WANG Jian2,LI Sheng-bin1 1.Forensic Laboratory of Ministry of Health of Xi'an Jiaotong University,710061,Xi'an China; 2.Beijing Huada Genomics Institute( Beijing Airport Industrial Zone B-6),101300,Beijing,China Abstract:In order to enrich the Chinese genetic database,nine polymorphic loci of STR,such as D3S1358,vWA,FGA,TH01,TPOX,CSF1PO,D5S818,D13S317 and D7S820 were studied.Based on STR gene scan marked by fluorescence,91 unrelated Chinese Chaoxian individuals were observed.81 alleles and 196 genotypes were found.The corresponding gene frequency and genotype frequency were 0.0055~0.4615 and 0.0110~0.9890 respectively.The genogypes frequency of nine STR loci was good with the Hardy-Weinberg equilibrium（P＞0.05）.The statistical analysis of nine STR loci showed the following：PIC（polymorphic information content）≥0.6863,H（heterozygosity）≥0.6919,DP（discrimination power）≥0.8301,EPP（probability of paternity exclusion）≥0.8590.The data studied can be used in Chinese population genetic studies and forensic medicine applications. Key words：Chaoxian groups of China;STRs;gene scan;genetic polymorphism     

广西环江县毛南族九个短串联重复序列基因座的遗传学分析

为了了解广西环江毛南族人群无关个体的九个短串联重复序列:vWA,D18S51,D5S818,FGA,D8S1179,D21S11,D7S820,D3S1358,D13S317基因座的遗传多态性分布情况;本文用枸橼酸钠抗凝法采集广西环江县毛南族200份无亲缘关系的健康个体的血样,Chelex-100方法提取DNA,应用AmpFlSTRIdentifilerTM荧光标记复合扩增技术对血样DNA的九个STR基因座进行扩增,用ABI 3100型遗传分析仪对扩增产物进行检测。结果显示九个STR位点的基因型分布均符合Hardy-Weinberg平衡定律,累积非父排除率达0.999996,累积个体识别能力达0.99999999996,多态信息总量为0.9999985。结论:广西环江县毛南族人群有自身的STR等位基因分布特征,所获数据可为法医学个体识别、亲子鉴定及群体的遗传学研究提供依据。     

用毛细管电泳技术对北京汉族人群9个STR基因座遗传多态性的研究

为对北京汉族D3S1358、vWA、FGA、D8S1179、D21S11、D18S51、D5S818、D13S317及D7S820等9个STR基因座的遗传多态性进行群体遗传学研究,利用荧光标记复合扩增及毛细管电泳自动荧光检测的方法,对236名无关个体获得9个STR基因座等位基因的分布频率,结果均符合Hardy-Weinberg平衡.计算了各基因座的杂合度（H）、个人识别能力（DP）、偶合率（PM）、非父排除率（EPP）和多态性信息总量（PIC）等群体遗传学数据.结果表明,这9个STR基因座多态性好,灵敏度高,可用于人类遗传分析及法医学中的亲子鉴定和个人识别.     

甘肃地区回族人群18个STR基因座的遗传多态性

研究了D5S818、D8S1179、D7S820、CSF1PO、D2S1338、D3S1358、v WA、D21S11、D16S539、Penta E、TPOX、TH01、D19S433、D18S51、FGA、D6S1043、D13S317、D12S391等18个短串联重复序列(short tandem repeats,STR)基因座在甘肃地区回族人群的遗传多态性。采用荧光标记复合扩增及毛细管电泳技术对1 038名甘肃地区回族无关个体18个STR基因座进行分析。研究结果显示,1 038名甘肃地区回族个体在18个STR基因座上,共检出223种等位基因,982种基因型,其分布均符合Hardy-Weinberg平衡(P0.05),18个基因座的杂合度(H)介于0.601~0.929之间,匹配概率(Pm)介于0.012~0.213之间,个体识别概率(DP)介于0.787~0.988之间,多态信息含量(PIC)介于0.550~0.920之间,非父排除概率(PE)值介于0.292~0.854之间。本文研究结果对甘肃地区回族人群群体遗传学及法医学后续研究应用具有参考价值。     

三个STR位点在哈萨克族中的遗传多态性

运用复合PCR扩增,6％变性聚丙烯酰胺凝胶电泳结合银染技术对我国新疆102位无关的哈萨克族个体进行D16S539,D7S820,D13S317的STR位点的调查,为建立新疆哈萨克族群体数据库提供资料。经统计学检验,3个位点的基因型频率分布符合Hardy-Weinberg平衡定律,结果显示3个位点的期望杂合度为：0.9439、0.9356、0.9304,累积PIC＝0.9905,DP＝0.9998,PE=9572。紫外,比较新疆哈萨克族与其他4个人群的等位片段频率,发现除与北京汉族在D7S820位点上无统计学意义外（P＞0.05）,其他均可见显著性差异（P＜0.05）。同时,在8个家系42人的调查中无一突变发现且均按孟德尔遗传规律传递。3个STR位点的联合分析在法医学应用及群体遗传学中显示了较高的价值。     

银染法检测短串联重复序列(STR)位点PCR产物鉴别MRC-5细胞

通过采用银染法鉴别短串联重复序列聚合酶链式反应(STR)位点的PCR产物来鉴别人二倍体细胞MRC-5株主细胞库、工作细胞库及限制代细胞。运用PCR方法对细胞库的9个STR位点(CSF1PO、TPOX、TH01、F13A01、FESFPS、vWA、D16S539、D7S820、D13S317)和性别鉴别位点Am elogen in进行扩增,变性聚丙烯酰胺凝胶电泳分离,银染法显影技术,检测MRC-5主细胞库、工作细胞库及限制代细胞的遗传标记。与ATCC公布的MRC-5的荧光STR图谱的8个STR位点和Am elogen in位点相比,MRC-5主细胞库、工作细胞库、限制代细胞的银染STR图谱的9个STR位点和Am elogen in位点,荧光法和银染法重叠的8个位点(CSF1PO、TPOX、TH01、FESFPS、vWA、D16S539、D7S820、D13S317和Am elogen in)数据完全吻合,说明细胞鉴别试验成立。STR图谱作为细胞鉴别的方法简单、易行、准确。     

Regeneration of multiple shoots from transgenic potato events facilitates the recovery of phenotypically normal lines: assessing a cry9Aa2 gene conferring insect resistance

Background

Tools for authenticating cell lines are critical for quality control in cell-based biological experiments. Currently there are methods to authenticate human cell lines using short tandem repeat (STR) markers based on the technology and procedures successfully used in the forensic community for human identification, but there are no STR based methods for authenticating nonhuman cell lines to date. There is significant homology between the human and vervet monkey genome and we utilized these similarities to design the first multiplex assay based on human STR markers for vervet cell line identification.

Results

The following STR markers were incorporated into the vervet multiplex PCR assay: D17S1304, D5S1467, D19S245, D1S518, D8S1106, D4S2408, D6S1017, and DYS389. The eight markers were successful in uniquely identifying sixty-two vervet monkey DNA samples and confirmed that Vero76 cells and COS-7 cells were derived from Vero and CV-1 cells, respectively. The multiplex assay shows specificity for vervet DNA within the determined allele range for vervet monkeys; however, the primers will also amplify human DNA for each marker resulting in amplicons outside the vervet allele range in several of the loci. The STR markers showed genetic stability in over sixty-nine passages of Vero cells, suggesting low mutation rates in the targeted STR sequences in the Vero cell line.

Conclusions

A functional vervet multiplex assay consisting of eight human STR markers with heterozygosity values ranging from 0.53-0.79 was successful in uniquely identifying sixty-two vervet monkey samples. The probability of a random match using these eight markers between any two vervet samples is approximately 1 in 1.9 million. While authenticating a vervet cell line, the multiplex assay may also be a useful indicator for human cell line contamination since the assay is based on human STR markers.     

奶牛和肉牛6个STR基因座遗传多态性研究

利用PCR技术和复合电泳银染技术检测奶牛和肉牛BM2113、BM1862、BMc701、BM2934、TGLA122、BM720等6个STR基因座的多态性分布,并计算该6个基因座的基因频率(P_i)、个体鉴别力(DP)、杂合度(H)、多态信息含量(PIC)、和非父排除概率(PE)。结果显示:6个STR基因座的基因型分布符合Hardy-Weinberg平衡,奶牛中6个STR基因座中BM2113基因座的DP、H和PIC最高,TGLA122基因座的PE最高。6个STR基因座的累积个体鉴别力(CDP)为0.99997,累积非父排除能力(CPE)为0.98827。肉牛中6个STR基因座中BM1862基因座的DP、H、PIC、PE都是最高,6个STR基因座的累积个体鉴别力(CDP)为0.99999,累积非父排除能力(CPE)为0.99578。结果表明,6个STR基因座可用于牛的遗传连锁分析、个体识别和亲子鉴定等研究领域。     

Paternity testing and forensic DNA typing by multiplex STR analysis using ABI PRISM 310 Genetic Analyzer

Short tandem repeats (STRs) are widespread throughout the human genome and are a rich source of highly polymorphic markers which can be detected by PCR. To gain a better appreciation for how the polymorphism at a particular locus impacts the individual identity, the present study was undertaken to explore the use of 15 STR loci in forensic investigation and paternity testing. Multiplex STR typing was used to study the 15 STR loci (D8S1179, D21S11, D7S820, CSF1PO, D3S1358, TH01, D13S317, D16S539, D2S1338, D19S433, vWA, TPOX, D18S51, D5S818 and FGA) in addition to a gender identification marker, amelogenin, by capillary electrophoresis on 310 Genetic Analyzer. Samples from 85 trio and duo cases of disputed paternity were investigated. The data were analyzed to give information on paternity index, probability of paternity, frequency of number of exclusions and rate of mismatch at each STR locus. The method was also successfully applied to forensic personal identification in theft and murder cases. The results demonstrated that the STR typing is a reliable and robust tool for analyzing the forensic practice as well as for paternity testing. The advantages of using multiplex STR analysis over other conventional methods are discussed.     

西藏那曲地区藏族人群15个短串联重复序列位点多态性的研究

利用基因扫描技术调查西藏自治区那曲地区藏族人群D8S1179、D21S11、D7S820、CSF1PO、D3S1358、TH01、D13S317、D16S539、D2S1338、D19S433、VWA、TPOX、D18S51、D5S818及FGA共15个短串联重复序列(STR)基因座多态性分布,获得15个基因座的群体遗传学数据。结果显示:15个STR位点在那曲地区藏族人群中具有遗传多态性,基因型分布符合Hardy-Weinberg平衡,DP在0.758 8—0.960 4之间,H在0.476 2—0.862 0之间,PIC在0.446 4—0.861 5之间,EPP在0.385 0—0.856 0之间,累积个体鉴别力为0.999 999 999,累积非父排除率为0.999 999 998。15个STR位点适合作为那曲地区藏族人群的遗传标记用于人类学、疾病连锁分析、法医学亲子鉴定和个体识别等领域的研究。     

Genetic data provided by 21 autosomal STR loci from Chinese Tujia ethnic group

The aim of this study was to investigate allelic frequency distribution and forensic genetic parameters of autosomal short tandem repeats (STR) loci of the population samples from 107 Tujia individuals from Chinese Hubei Province. Twenty-one autosomal STR genetic markers (D9S1122, D6S474, D6S1017, D5S2500, D4S2408, D3S4529, D2S441, D2S1776, D22S1045, D20S482, D1S1677, D1S1627, D1GATA113, D19S433, D18S853, D17S1301, D11S4463, D12ATA63, D10S1248, D10S1435 and D14S1434) were simultaneously amplified in a new multiplex polymerase chain reaction system. 155 alleles for all the STR loci from the Tujia population were observed and the corresponding allelic frequencies ranged from 0.005 to 0.589. Expected heterozygosity, polymorphic information content, power of discrimination and power of exclusion of the 21 STR loci in the Tujia population were from 0.579 to 0.824, from 0.525 to 0.802, from 0.773 to 0.945 and from 0.257 to 0.641, respectively. Our results indicate that the autosomal STRs multiplex system provides highly informative STR data and could be useful in forensic individual identification and parentage testing in this region.     

西藏藏族人群15个短串联重复序列基因座的遗传多态性

利用多重PCR五色荧光(6FAM、VIC、NED、PET、LIZ)自动化检测技术检测西藏自治区藏族人群D8S1179、D21S11、D7S820、CSF1PO、D3S1358、TH01、D13S317、D16S539、D2S1338、D19S433、VWA、TPOX、D18S51、D5S818及FGA共15个STR基因座遗传多态性, 获得15个STR基因座的群体遗传学数据。结果显示：15个STR基因座的基因型分布符合Hardy-Weinberg平衡。15个STR基因座的个体鉴别力 (Discrimination power, DP)在0.7555~0.9602之间, 杂合度 (Heterozygosity, H)在0.5651~0.8530之间, 多态性信息含量 (Polymorphism information content, PIC)在0.5528~0.8456之间, 非父排除率(Probability of paternity exclusion, EPP)在0.3811~0.8549之间, 累积个体鉴别力为0.999999999, 累积非父排除率为0.999999998。15个短串联重复序列基因座适合作为西藏藏族人群的遗传标记, 用于人类学、疾病连锁分析、法医学亲子鉴定和个体识别等领域的研究。     

Two-color multiplexed analysis of eight short tandem repeat loci with an electrophoretic microdevice.

Single-channel microfabricated electrophoretic devices equipped with a dual-wavelength laser-induced fluorescence detection system were used for the fast analysis of an eight-loci, two-color multiplex short tandem repeat (STR) system for human identification. Routine analyses of the eight loci (CSF1PO, TPOX, TH01, vWA and D16S539, D7S820, D13S317, D5S818), requiring four-base resolution, were performed in only 2 min. Specific analyses for a microvariant allele (allele 9.3 of the TH01 locus) demanded single-base resolution and was performed in less than 10 min. The high accuracy of the microdevice for real-world STR sample analyses was demonstrated by comparison with conventional slab-gel electrophoresis. Our results show that a fast multiwavelength multichannel electrophoretic microsystem will be capable of routinely processing thousands of complex STR samples per day.