单核苷酸多态性技术在鸡遗传变异中的研究及应用

单核苷酸多态性(SNP)是继限制性片段长度多态性、微卫星标记之后的第三代分子标记,通常呈双等位基因多态。本文从SNP的特点、SNP的发现与检测、SNP数据库、SNP的频率及其与表型的关系等方面简述了SNP在鸡遗传变异中的研究及应用进展。     

单核苷酸多态性及其在鸡QTL定位上的应用

单核苷酸多态性是指DNA序列上的单个碱基变异,它具有分布广、多态信息含量大、易于检测和统计分析等优点,能较好用于基因图谱构建和数量性状QTL定位研究,被称为继RFLP和微卫星标记之后的第3代基因遗传标记。本文综述了单核苷酸多态性的性质及检测技术、利用候选基因SNP进行鸡QTL定位研究的现状,并对未来SNP的应用前景进行了展望。Abstract:Single nucleotide polymorphism (SNP) refers to the change of single nucleotide in DNA sequence.Because of its high density in genomes and easy in detection and analysis statistically,SNP can be used in genetic linkage map construction and QTL mapping.Here,the characters and detecting technology of SNP,as well as the status and foreground of the use of candidate gene SNP in chicken QTL mapping are introduced.     

单核苷酸多态性及其在水稻中的应用(综述)

单核苷酸多态性(single nucleotide polymorphism,SNP)主要是指基因组的DNA由于单个核苷酸的变异所引起的DNA序列多态性。本文介绍SNP的检测方法及其在水稻中的研究进展和应用。     

毒蛇咬伤病人的血小板聚集功能初探

目的为了探讨血液毒及混合毒类毒蛇咬伤病人的血小板功能情况,了解临床中毒发病机理。方法对近年来在本院急诊科确诊的五步蛇(Da.),蝮蛇(Ah.),眼镜蛇(Nn.),眼镜王蛇(Oh.),蝰蛇(Vr.),烙铁头(Tm.)和竹叶青(Ts.)等各种血液毒及混合毒类毒蛇咬伤17例病人25个血样,进行最大血小板聚集率(MAR)的检查和分析研究。结果除了Da未见MAR下降外(仅1例1个样品),其余各蛇种咬伤的样品均有下降。在获得未经治疗(抗蛇毒血清)的15个样品的检查结果中,进行统计分析,Vr、Tm以及Ah的MAR比正常人对照组(51.3±16.0)%明显下降。结论蛇毒中的血小板集聚抑制因子,可引起蛇伤患者体内血小板聚集功能下降,血小板处于无用状态,结果有些蛇伤病人DIC样综合征时出现严重出血而无明显微循环血栓形成致器官受损的临床症状。     

单核苷酸多态性在作物遗传及改良中的应用

单核苷酸多态性（single nucleotide polymorphism,SNP）是等位基因间序列差异最为普遍的类型,可作为一种高通量的遗传标记。已建立了PCR扩增目标序列及其产物测序和电子SNP（eSNP）等多种发现和检测SNP的方法。玉米和大豆等作物也已开展了SNP分析。一些栽培作物种质的多样性不断减少,其结果使连锁不平衡（linkage disequilibrium,LD）增加,这有利于目的基因座上SNP单元型（haplotype）与表型的相关性分析。SNP已在作物基因作图及其整合、分子标记辅助育种和功能基因组学等领域展示了广泛的应用价值。 Abstract:Single nucleotide polymorphism(SNP) is the most common type of sequence difference between alleles,which can be used as a kind of high-throughput genetic marker.Several different routes have been developed to discover and identify SNP.These include the direct sequencing of PCR amplicons,electronic SNP(eSNP) and so on.SNP assays have been made in many crop species such as maize and soybean.The elite germplasm of some crops have been narrowed in genetic diversity,increasing the amount of linkage disequilibrium(LD) present and facilitating the association of SNP haplotypes at candidate gene loci with phenotypes.SNP analysis has been broadly used in the field of plant gene mapping,integration of genetic and physical maps,DNA marker-assisted breeding and functional genomics.     

单核苷酸多态性检测技术在法医学中的应用进展

单核苷酸多态性是新一代的法医学检测标记,本文结合市面上常见的商品化试剂盒,对单核苷酸多态性的检测原理、试剂选择及具体操作等进行了综述.并在此基础上提出了它们在法医学应用中需要注意的问题.     

CYP1A1基因单核苷酸多态性与肺癌的相关性研究

目的:探讨代谢酶CYP1A1基因MspI位点多态性与新疆汉族人群肺癌遗传易感性之间的相关性.方法:应用聚合酶链式反应(PCR)-限制性片段长度多态性(RFLP)技术检测59例新疆汉族肺癌和84例新疆汉族健康人的CYP1A1基因MspI位点多态性分布频率,并分析了CYP1A1基因MspI位点多态性与新疆汉族人群肺癌遗传易感性和患者性别之间的相关性.结果:(1)CYP1A1基因MspI位点3种多态基因型分布频率在两组间比较差异有统计学意义(χ2=6.682,P=0.035),CC基因型在病例组的分布频率显著高于正常对照组.(2)携带突变CC基因型的个体较携带TT基因型的个体患肺癌的危险性增加(OR=3.759.95%CI=1.228-11.494,P=0.035).(3)男女肺癌患者的CYP1A1基因MspI位点基因型及等位基因频率的差异均无显著性(P>0.05).结论:(1)CC突变基因型可能是新疆汉族人群的肺癌易感因素.(2)CYP1A1基因MspI位点多态性可能与新疆汉族肺癌患者的性别无关.     

Molecular Cloning and Single Nucleotide Polymorphism Detection of Buffalo DGAT1 Gene

We report molecular cloning and single nucleotide polymorphism detection of the buffalo DGAT1 gene. Diacylglycerol acyltransferase (DGAT1) is considered the key enzyme in controlling the rate of synthesis of triglycerides. The DGAT1 gene was recently identified as a strong functional candidate gene affecting milk yield and composition in cattle. A full-length buffalo DGAT1 genomic DNA was amplified by iterative PCR based on homolog cloning. The buffalo DGAT1 gene comprises 17 exons and spans approximately 8.3 kb. The genomic structures of DGAT1 are highly conserved among mammal species. The deduced protein of buffalo DGAT1 contains 489 amino acids, showing high-sequence similarity with mammal homologs. Through PCR-SSCP analysis and sequencing, seven polymorphic positions were detected in the complete genomic region of buffalo DGAT1, and their frequencies were observed from a collection of 117 buffalo. The SNP (C/T) detected at position 11785 in exon 17 creates a substitution change for the amino acid sequence, resulting in an Ala residue (GCG) transition to a Val residue (GTG) in position 484 of buffalo DGAT1 protein. Information provided in this study will be useful in further studies to determine the role DGAT1 plays in the regulation of milk fat synthesis and quality improvement for milk in buffalo. Jing Yuan and Jun Zhou contributed equally to this work.     

水稻单核苷酸多态性及其应用现状

单核苷酸多态性（single nucleotide polymorphisms, SNPs）在水稻中数量多,分布密度高,遗传稳定性高。水稻SNPs的发现方法主要有对样本DNA的PCR产物直接测序、从SSR区段检测SNPs和从基因组序列直接搜索等。目前已有多种基因分型技术运用到了水稻SNPs检测,SNPs检测的高度自动化使水稻SNPs基因分型非常方便。单核苷酸多态性在水稻遗传图谱的构建、基因克隆和功能基因组学研究、标记辅助选择育种、遗传资源分类及物种进化等方面的应用具有巨大潜力。     

猪Kappa阿片受体基因外显子部分序列单核苷酸多态性分析

应用PCR SSCP的方法对大白猪、长白猪和杜洛克猪的Kappa阿片受体 (Kappaopioidreceptor,简称KOR)基因进行单核苷酸多态性检测和分析 ,研究Kappa阿片受体基因作为候选基因 ,影响母猪行为规癖性状的可能性。根据Kappa阿片受体基因外显子的部分序列设计 3对引物 ,发现F1/R1引物对扩增的片段有多态性。对两种纯合子片段克隆并测序表明 ,mRNA第 10 5处有一C→T的单碱基突变 ,为沉默突变。统计结果发现 ,3种基因型 (AA、AB、BB)在各品种中的分布不一致 ,χ2 独立性检验差异极显著 (P <0 0 1)。将 3种基因型同行为规癖性状进行统计分析 ,结果表明BB基因型与其他两种基因型相比表现较高的静止站立行为 ,同AA和AB型比较差异极显著 (P<0 0 1) ,其他 4种行为性状基因型间差异不显著。因此 ,推测Kappa阿片受体基因SNP对母猪静止站立行为性状存在一定的影响     

中国广东汉族人群核苷酸修复基因hMTH1 遗传多态性研究

为研究中国南方汉族人群核苷酸修复基因hMTH1遗传多态性,应用聚合酶链反应-单链构象多态性技术检测172名健康人外周血白细胞hMTH1基因启动子及全部5个外显子多态性,并进行DNA测序。结果发现hMTH1基因启动子及外显子1序列保守,未见突变;外显子2第73位碱基存在T→C杂合型突变,基因型TT和TC频率分别为93.02%、6.98%,等位基因T和C频率分别为96.51%、3.49％;外显子3第45位遗传密码存在T→C杂合型突变,基因型TT和TC频率分别为95.35%、4.65%,等位基因T和C频率分别为97.67%、2.33%,该多态性为首次发现;外显子4第83位遗传密码存在G→A杂合型突变,基因型GG和GA频率分别为89.53%、10.47%,等位基因G和A频率分别为94.77%、5.23％;外显子5第119位氨基酸遗传密码存在C→T杂合型突变,基因型CC和CT频率分别为95.93%、4.07%,等位基因C和T频率分别为97.97%、2.03％。Abstract: In order to study the genetic polymorphisms of nucleotide repair gene hMTH1 in southern Chinese Han population, the polymorphisms of the gene’s promoter and its five exons among peripheral blood lymphocytes of 172 Chinese Han people were analyzed with polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) and DNA sequencing. The sequences of the promoter and exon 1 of hMTH1 gene were conserved. A T to C polymorphism was detected at the 73th base in exon2. The genotype frequencies of TT and TC were 93.02% and 6.98%, respectively. The allelic frequencies of T and C were 96.51% and 3.49％, respectively. A T to C polymorphism was detected at codon 45 in exon3, which was first reported. The genotype frequencies of TT and TC were 95.35% and 4.65%, respectively. The allelic frequencies of T and C were 97.67% and 2.33%, respectively. A G to A polymorphism was detected at codon 83 in exon4. The genotype frequencies of GG and GA were 89.53% and 10.47%, respectively. The allelic frequencies of G and A were 94.77% and 5.23％, respectively. A C to T polymorphism was detected at codon 119 in exon5. The genotype frequencies of CC and CT were 95.93% and 4.07%, respectively. The allelic frequencies of C and T were 97.97% and 2.03％, respectively.     

ABCB1 基因单核苷酸多态性rs2235048 与氯吡格雷抵抗的相关关系

目的:探讨ABCB1基因单核苷酸多态位点rs2235048在中国汉族人群中的分布及其与氯吡格雷抵抗(Clopidogrelresistance,CR)发生的相关关系。方法:采用光学比浊法测定20μmol/L ADP诱导的残余血小板聚集率(Residual plateletagglutination,RPA)。当RPA≥70%时,即为CR。所有入选患者分为CR组和氯吡格雷非抵抗组(Non-clopidogrel resistance,NCR)。采用焦磷酸测序法测定ABCB1基因rs2235048单核苷酸多态位点在CR组和NCR组的基因型及等位基因分布频率。结果:ABCB1基因rs2235048多态在CR组和NCR组基因型分布频率符合Hardy-Weinberg平衡。在CR组和NCR组中,ABCB1基因rs2235048多态的基因型频率分布无统计学差异(P=0.527,X2=1.281);T、C等位基因频率在两组间分布频率也没有统计学差异(P=0.740,OR=0.958,95%CI=0.742～1.236)。结论:对PCI术后服用氯吡格雷的冠心病患者进行分析发现,ABCB1基因单核苷酸多态位点rs2235048与冠心病患者CR的发生无相关关系。     

替格瑞洛与氯吡格雷对急性冠脉综合征患者经皮冠脉动脉介入术后血小板抑制效果的比较

目的:比较替格瑞洛与氯吡格雷对急性冠脉综合征(ACS)患者经皮冠脉动脉介入术(PCI)后血小板的抑制效果。方法:选择2014年3月至8月在我院经替格瑞洛联合阿司匹林治疗的ACS患者85例(替格瑞洛组),按性别、年龄2:1匹配原则随机抽取同一时间服用氯吡格雷联合阿司匹林治疗患者170例(氯吡格雷组)为研究对象,两组患者均行PCI治疗,并于服用抗血小板药物负荷剂量2天(PCI术后)进行血栓弹力图(TEG)检测,观察和比较两组患者经ADP途径及经AA途径的血小板抑制率。结果:氯吡格雷组和替格瑞洛组经ADP途径的血小板抑制率分别为(66.60±25.57)%、(82.10±18.87)%,两组比较差异有统计学意义(P0.05)。氯吡格雷组ADP抑制率50%患者占总人数的29.4%,替格瑞洛组ADP抑制率50%的患者占总人数的10.6%,两组差异有统计学意义(P0.05),氯吡格雷组ADP抑制率75%者占总人数的41.8%;而替格瑞洛组ADP抑制率75%的患者占总人数的69.4%,两组抑制率差异也存在统计学意义(P0.05)。氯吡格雷组和替格瑞洛组经AA途径的血小板抑制率分别为(88.70±23.89)%、(90.32±18.09)%,两组比较差异无统计学意义(P0.05)。结论:替格瑞洛对ACS患者PCI术后血小板的抑制作用优于氯吡格雷。     

A Single Nucleotide Polymorphism and Sequence Analysis of CSN1S1 Gene Promoter Region in Chinese BOS Grunniens (YAK)

The aim of this study was to investigate the polymorphism of the CSN1S1 gene promoter region in 4 Chinese yak breeds, and compare the yak CSN1S1 gene promoter region sequences with other ruminants. A Polymerase Chain Reaction-Single Strand Conformation Polymorphism protocol was developed for rapid genotyping of the yak CSN1S1 gene. One hundred fifty-eight animals from 4 Chinese yak breeds were genotyped at the CSN1S1 locus using the protocol developed. A single nucleotide polymorphism of the CSN1S1 gene promoter region has been identified in all yak breeds investigated. The polymorphism consists of a single nucleotide substitution G→A at position 386 of the CSN1S1 gene promoter region, resulting in two alleles named, respectively, G₃₈₆ and A₃₈₆, based on the nucleotide at position 386. The allele G₃₈₆ was found to be more common in the animals investigated. The corresponding nucleotide sequences in GenBank of yak (having the same nucleotides as allele G₃₈₆ in this study), bovine, water buffalo, sheep, and goat had similarity of 99.68%, 99.35%, 97.42%, 95.14%, and 94.19%, respectively, with the yak allele A_386.     

血清镁、血小板聚集率在糖耐量减低患者微血管病变中的临床意义

目的:观察糖耐量减低患者微血管病变与血清镁、血小板聚集率(PAR)的相关性。方法:将2012年11月-2014年1月在本院内分泌科和体检中心的120例研究对象随机均分为糖耐量减低合并微血管病变组(n=30),糖耐量减低无微血管病变组(n=30),糖尿病组(n=30),正常对照组(n=30)。比较各组患者的生化指标、血清镁与PAR的相关性、血镁水平以及PAR与FPG、HbA1C相关性。结果:糖尿病组和糖耐量减低合并微血管病变组患者的所有生化指标与对照组相比,差异有显著性(P0.05);血清镁水平与PAR之间呈显著负相关(P0.01);血镁水平与糖耐量减低合并微血管病变组、糖耐量减低无微血管病变组、糖尿病组中FPG、HbA1C呈负相关关系(P0.05);PAR与糖耐量减低合并微血管病变组、糖耐量减低无微血管病变组、糖尿病组中FPG、HbA1C呈正相关关系(P0.05)。结论:血清镁与PAR在预测糖耐量患者微血管病变中有一定的临床意义,值得临床推广。     

Polymorphism of the SOD1-DNA aggregation species can be modulated by DNA

Proteinaceous aggregates rich in copper, zinc superoxide dismutase (SOD1) have been found in both in vivo and in vitro models. We have shown that double-stranded DNA that acts as a template accelerates the in vitro formation of wild-type SOD1 aggregates. Here, we examined the polymorphism of templated-SOD1 aggregates generated in vitro upon association with DNA under different conditions. Electron microscopy imaging indicates that this polymorphism is capable of being manipulated by the shapes, structures, and doses of the DNAs tested. The nanometer- and micrometer-scale aggregates formed under acidic conditions and under neutral conditions containing ascorbate fall into three classes: aggregate monomers, oligomeric aggregates, and macroaggregates. The aggregate monomers observed at given DNA doses exhibit a polymorphism that is markedly corresponded to the coiled shapes of linear DNA and structures of plasmid DNA. On the other hand, the regularly branched structures observed under both atomic force microscopy and optical microscope indicate that the DNAs tested are simultaneously condensed into a nanoparticle with a specific morphology during SOD1 aggregation, revealing that SOD1 aggregation and DNA condensation are two concurrent phenomena. The results might provide the basis of therapeutic approaches to suppress the formation of toxic protein oligomers or aggregates by screening the toxicity of the protein aggregates with various sizes and morphologies.     

中国汉族人群MASP2基因单核苷酸多态性的连锁不平衡和单体型分析

目的：探讨中国汉族人群甘露糖结合凝集素相关的丝氨酸蛋白酶（MASP2）基因单核苷酸多态性（SNP）的连锁不平衡和单体型,为相关研究提供更为详实的数据。方法：选取30例广州汉族无关个体,用重新测序的方法进行MASP2基因SNP的发掘,构建连锁不平衡（LD）模式,与HapMap公布的其他4个人群数据相比,并选择4个标签SNP（tagSNP）在232例北京汉族个体和291例广州汉族个体中分析MASP2的多态性和单体型。结果和结论：对MASP2的重测序共检测出16个SNP,LD分析显示来自中国不同地区人群的LD模式基本相同,与来自美国和日本的人群存在差异;北京和广州地区汉族人群tagSNP的等位基因和基因型频率分布不存在显著差异。     

单核苷酸多态性及其在油菜分子标记辅助育种中的应用

油菜是一种重要的油料作物,油菜的基因组功能研究已引起了各国政府和科研工作者的高度重视。SNP是目前为止分布最广泛,存在数量最多且标记密度最高的一种遗传多态性标记。对油菜等作物的基因组功能研究提供了丰富的信息资源。综述了近来单核苷酸多态性的研究进展以及其在油菜的分子标记辅助品质育种中的可能用途。     

牛FSHR基因第10外显子单核苷酸多态性及其与双胎性状的关系

以秦川牛和荷斯坦奶牛的双胎母牛和单胎母牛为实验材料 ,以牛的FSHR基因的第 10个外显子作为标记牛双胎性状的候选基因 ,用SNP法进行了多态检测 .结果发现 ,在秦川牛的双胎母牛中突变率 6 0 % (6 10 ) ,而在单胎母牛中突变率为 2 0 % (2 10 ) ;在荷斯坦奶牛中 ,双胎母牛突变率为31 2 5 % (5 16 ) ,单胎母牛突变率为 6 6 7% (1 15 ) ;由此可见双胎牛和单胎牛二者之间FSHR基因的第 10个外显子的突变率差异明显 .这表明 ,选择FSHR基因的第 10个外显子有可能作为双胎性状的候选基因 .序列分析发现 ,在FSHR基因的第 15 0 6位碱基发生了突变 (T→C) ,但氨基酸没有发生变化 .     

鸡瘦蛋白受体(OBR)基因内含子8单核苷酸多态性与体脂性状的相关研究

以东北农业大学高低脂双向选择系的第 6世代肉鸡为材料 ,鸡 7周龄时测定体重和腹脂重等屠体性状。根据鸡瘦蛋白受体基因内含子 8的序列 (GenBank登陆号 :AF2 2 2 783 )设计引物 ,用直接测序的方法检测多态性位点 ,用PCR SSCP的方法进行基因型分析 ,建立适合的统计模型对多态性位点产生的基因型与生长和体组成性状进行相关分析。结果表明 ,在第 50 0和 659位碱基同时发生了T—C、G—A突变。经最小二乘分析 ,3种基因型在腹脂重和腹脂率上差异显著 (P <0 0 5) ,BB型个体腹脂重和腹脂率显著高于AB型 (P <0 0 5) ,极显著地高于AA型个体 (P <0 0 1) ;3种基因型在肝重上差异显著 (P <0 0 5) ,且AA基因型个体的肝重显著低于AB和BB基因型个体。初步推断OBR基因可能是影响鸡脂肪性状的主效基因或与主效基因连锁 ,推测可以利用这个多态位点对鸡的体脂性状进行标记辅助选择