云南6个地区恒河猴蛋白多态及遗传多样性研究

云南６个地区恒河猴蛋白多态及遗传多样性研究＊ＧＥＮＥＴＩＣＤＩＶＥＲＳＩＴＹＡＭＯＮＧＭａｃａｃａｍｕｌａｔａＦＲＯＭＳＩＸＲＥＧＩＯＮＳＩＮＹＵＮＮＡＮＰＲＯＶＩＮＣＥＢＡＳＥＤＯＮＰＲＯＴＥＩＮＥＬＥＣＴＲＯＰＨＯＲＥＳＩＳ关键词恒河猴，蛋白电泳...     

恒河猴与食蟹猴微卫星DNA多态性的比较分析

目的分析恒河猴和食蟹猴群体间的遗传多样性,确立一种对恒河猴和食蟹猴种群个体的遗传鉴别方法。方法利用聚合酶链反应（PCR）扩增技术采用15个多态性微卫星DNA位点对50只恒河猴和50只食蟹猴个体进行了DNA多态性的分析,对比两群体间等位基因数目差异。结果筛选的15个具有显著多态性的微卫星DNA位点对恒河猴和食蟹猴种群可以进行DNA多态性分析,其等位基因数目均在7个以上,且两群体间有11个位点的等位基因数存在一定的差异。结论利用这些多态性微卫星DNA位点建立一种有效鉴别恒河猴和食蟹猴种群遗传背景的方法具有一定的可行性。     

中国驴种线粒体DNA D-loop多态性研究

利用Clustal W软件对我国5个家驴品种26个个体的mtDNA D—loop区399bp序列进行同源序列比对,共检测到核苷酸多态位点23个,只有转换1种类型,约占所测核苷酸的5．76％。以欧洲驴D—loop作对照,我国5个家驴品种D—loop区序列的平均核苷酸变异率为1．80％,其中凉州驴的平均核苷酸变异率为0．35％．云南驴为1．25％,关中驴为2．30％,新疆驴为2．91％,佳米驴为2．20％。家驴品种内与品种间mtDNA D—loop区序列歧异度分别为0．25％-5．01％和4．51％-5．51％,说明家驴品种间D—loop区序列多态性比较丰富。在所测家驴个体中,mtD NAD—loop序列由11种单倍型组成,单倍型比例为42．31％,表明我国家驴mtDNA遗传多态性正逐步丧失,需要加强其种质资源保护。引用GenBank中亚洲野驴和欧洲家驴的序列,构建了我国5个家驴品种的NJ分子系统树,首次从分子水平证实中国家驴可能起源于非洲野驴,而与亚洲野驴无关。     

山东荣成人群线粒体DNA多态性研究

人类线粒体DNA（mtDNA)COⅡ/tRNA^lys区有两个9－bp(CCCCCTCTA)的串联重复序列,此重复序列中一个重复单位的缺失,在亚态地区人群中很普遍。对210名山东荣成人的mtDNA COⅡ/tRNA^lys区的9-bp 缺失情况进行了检测,并从中随机选取95个样本,利用PCR－RFLP法对另外6个区进行了多态性分析,以确定其单位型。结果表明,荣成人9－bp缺失频率为12.4%,相对于已检测的中国其他群体,此缺失频率处于中等水平。同时多态性分析也表明在95个被检测对象中存在27种不同的单倍型。此外还发现了两个未报道过的新酶切位点,序列分析表明是由点突变造成的。     

褐家鼠线粒体DNA遗传多态性的研究

通过碱变性法提取线粒体ＤＮＡ,用地高辛标记的探针Ｓｏｕｔｈｅｒｎ杂交限制性酶切多态性（ＲＦＬＰ）分析,研究中国家鼠Ｒａｔｔｕｓｎｏｒｖｅｇｉｃｕｓ遗传多态性。采用ＡｐａⅠ、ＡｖａⅠ、ＢａｎＨＩ、ＢｃｌⅠ、ＢｇｌⅠ、ＣｌａⅠ、ＥｃｏＲⅠ、ＥｃｏＲⅤ、ｈｉｎｄⅢ、ＰｖａⅡ、ＳｃａⅠ和ＸｂａⅠ等１２种限制性内切酶分析来自我国８个地区２６只褐家鼠的线粒体ＤＮＡ,共检出２０种限制性态型和１１种ｍｔＤＮＡ单倍     

普氏野马线粒体DNA D-loop区序列多态性研究

目的:了解中国新疆吉木萨尔野马繁殖中心的普氏野马(Equus przewalskii)遗传多样性及其遗传背景.方法:采用PCR产物直接测序法,对15匹普氏野马线粒体DNA D-loop高变区进行测序分析.结果:测定15个个体的线粒体DNA D-loop高变区15464～15866片段序列402bp.检测到12种单倍型,包括37个多态位点,占全部序列的9.2%,其中转换位点24个、颠换位点20个、转换位点和颠换并存位点8个、缺失位点3个.A%+T%含量(56.1%)高于G%+C%含量(43.9%),平均A含量为28.4%,T含量为27.7%,C含量为29%,G含为14.9%.单倍型间平均遗传距离为0.030,单倍型多态性(h)为1±0.00116,核苷酸多态性(π)为2.90%.15匹普氏野马线粒体DNA D-loop高变区之间平均核苷酸变异率为2.48%.结论:研究表明我国新疆吉木萨尔野马繁殖中心的普氏野马线粒体DNA D-loop区序列存在丰富的多态性.     

中国部分牦牛品种线粒体DNA遗传多态性研究

采用DNA测序技术首次测定了中国5个牦牛品种(类群)35个个体线粒体DNA控制区(D loop)全序列,结果表明:牦牛线粒体DNA控制区全序列长度为891～895bp,T、C、A、G4种核苷酸的含量分别为28.5%、25.3%、32.5%、13.7%。检测到55个变异位点,约占分析位点总数的6.16%,核甘酸变异类型有转换、颠换、插入/缺失4种。确定了24种单倍型,单倍型H4和H6为中国牦牛的主体单倍型,各单倍型在品种间分布不平衡,所测定的5个牦牛品种(类群)单倍型多样度平均为0.9697±0.0180,说明我国牦牛D loop单倍型类型丰富。牦牛品种内各序列平均核苷酸差异数为10.936,核苷酸多样度为1.231%;牦牛品种间核苷酸分歧度(Dxy)约为0.760%～2.155%,品种间双参数距离范围为0.000～0.029,表明我国牦牛遗传多态性丰富。对D环序列变异的分子方差分析和单倍型网络关系图的结果表明:我国牦牛品种间出现显著的遗传分化,牦牛单倍型网络关系图聚为2个聚类簇,表明我国牦牛有2个母系来源,或者有2个主要的驯化地点。     

恒河猴群微卫星DNA多态性的分析

目的　确立一种对恒河猴群个体的遗传物质进行准确可靠、快速简便的遗传检测方法。方法　利用聚合酶链反应 (PCR)扩增技术对 2 0只恒河猴群个体间进行了DNA多态性的分析。结果　筛选出 9个微卫星DNA位点具有显著多态性 ,4个微卫星DNA位点没有多态性 ,还有 2个位点等位基因数目较少。结论　利用这些多态性微卫星位点建立一种对恒河猴群个体进行有效、准备可靠、快捷简便的遗传背景监测方法。     

中国部分黄牛品种mtDNA遗传多态性研究

对我国8个黄牛品种22个个体的mtDNA D-loop区910bp全序列进行了分析。结果表明：8个黄牛品种D-loop区序列中,A T平均含量为61．65％;经比对,共检测到66个核苷酸多态位点,约占核苷酸总数的7．25％;D-loop全序列突变类型有5种,即转换、颠换、插入、缺失及转换与颠换共存,它们分别占核苷酸多态位点的81．82％、6．06％、7．57％、3．03％及1．52％。以欧洲牛mtDNA D-loop全序列为标准,8个黄牛群体D-loop的平均核苷酸变异率分3个层次：西镇牛、蒙古牛、黑白花牛及秦川牛的核苷酸变异率最低,分别为0．37％、0．44％、0．52％和0．66％;南阳牛与郏县红牛的核苷酸变异率居中,分别为1．91％和2．02％;晋南牛与岳阳牛的核苷酸变异率最高,分别为4．47％和4．73％。中国黄牛品种内D-loop区序列歧异度为0．55％～5．39％,品种间序列歧异度为1．21％～6．59％。在所测黄牛个体中,mtDNA D-loop序列由19种单倍型组成,单倍型比例为86．36%,说明中国黄牛mtDNA遗传多态性很丰富。由此构建了中国8个黄牛品种的NJ分子系统树,聚类分析表明：所测黄牛的mtDNA D-loop序列表现为3个单倍型组,从而揭示中国黄牛可能有3个母系起源,以普通牛起源和瘤牛起源为主。     

线粒体DNA多态性在海洋动物群体遗传结构研究中的应用

线粒体DNA( mtDNA)分析在揭示物种亲缘关系、遗传比较、系统进化和遗传结构等领域的研究中得到了广泛的应用,尤其是在海洋动物的遗传结构研究中发挥了重要的作用.介绍线粒体DNA的结构特征、多态性研究方法,并对其在海洋动物群体遗传结构研究中的应用进行了综述.     

云南猕猴mtDNA控制区全序列测定

目的测定云南猕猴线粒体DNA控制区全序列,对其进行鉴定及进化分析。方法利用PCR技术扩增猕猴线粒体DNA控制区全序列,结合GenBank中下载的猕猴参考序列（AY612638）,采用多个生物学软件对序列碱基组成、同源性、转换／颠换比等遗传信息进行分析,并基于邻接法（NJ）和最小进化法（ME）构建系统进化树。结果云南猕猴线粒体DNA控制区全长为（1084-1089）bp,A、T、G和c四种碱基平均含量分别为29．9％、26．9％、12．3％和30．9％,A＋T含量（56．8％）高于G＋C含量（43．2％）。所分析序列间的同源性为91．5％-99．5％,平均核苷酸变异率为4．5％,变异类型包括转换、颠换、插入和缺失4种形式,转换／颠换比值平均为26．1。进化树显示云南猕猴存在两个平行进化的姐妹分支。结论本研究获得了云南猕猴mtDNA控制区全序列,为猕猴进化关系研究及mtDNA控制区功能研究奠定基础。     

Genetic variation in transferrin alleles of rhesus macaques, Macaca mulatta

The rhesus macaque displays an extensive polymorphism at the transferrin locus. A principal components analysis describes the variance and covariance of alleles at the transferrin locus in eight widely dispersed sample populations. Using an eigenvectorial representation of the covariance matrix and systematically approximated geographical locations the distribution of populations and transferrin alleles is compared. Alleles with high variance prove to be the determining factor in the placement of populations in a "genetic map" and provide a means for interpreting the low congruence of genetics and geography found.     

Genetic studies of serum transferrins of free-ranging rhesus macaques of Cayo Santiago, Macaca mulatta (Zimmerman 1780)

Transferrin phenotypes of plasma from 687 semi-free-ranging Macaca mulatta living on Cayo Santiago, Puerto Rico, were determined by starch-gel electrophoresis and autoradiography. Fifteen phenotypes, homozygous or heterozygous products of six codominant autosomal alleles, were present in the population. The 687 animals, prior to March 1973, were divided into eight troops plus peripheral males. By March 1, 1973 the size of the population was reduced to 333 animals, consisting of four troops plus peripheral males. The distributions of transferrin phenotypes and allele frequencies were determined for the population of 687 animals and for 287 of the March 1973 population of 333 animals. Gametic ratios of 382 offspring of 126 females were enumerated. Statistical tests of homogeneity and equilibrium were applied to the data. The results of these tests suggest that, with only a few exceptions, the Cayo Santiago macaques, although divided into social groups, form a single population, and the results are in accord with behavioral observations. The authors suggest that the data on transferrins provide a good baseline for future genetic and ethological studies of evolutionary processes in a population of nonhuman primates.     

The albumin polymorphism and bilirubin binding in rhesus monkeys (Macaca mulatta)

Family studies confirmed a codominant mode of inheritance of albumin phenotypes identified in Macaca mulatta. Preliminary epidemiological and bilirubin binding studies suggest that selection favors the variant allele by reducing risks of hyperbilirubinemia and might be responsible for this otherwise rare polymorphism.     

Blood group antigens and the population genetics of Macaca mulatta on Cayo Santiago. I. Genetic differentiation of social groups

The red blood cell phenotypes for eight polymorphic loci were determined for 293 free-ranging Macaca mulatta living on Cayo Santiago; this number represents the total population of the island, disposed in four social groups plus peripheral males. The rhesus population shows significant genetic heterogeneity over blood group systems (loci) and social groups. No particular genetic locus or social group is solely responsible for the genetic heterogeneity observed. The distributions of genotypes for two loci (G and H) do not deviate significantly from Hardy-Weinberg expectations within social groups or in the population as a whole. Correction of the equilibrium expectations for the effect of population subdivision yields no statistically significant results. Overall, the results suggest that the interaction of a variety of processes (random genetic drift, founder effect, migration and selection) may be responsible for the diversity observed. These data, combined with those from further studies, may allow an application of behavioral and genetic knowledge to the study of microevolutionary processes among nonhuman primates.     

四川地区猕猴线粒体DNA 控制区遗传多样性及其种群遗传结构

猕猴是最理想的医学实验灵长类动物,并且是国家二级保护动物。四川地区的猕猴数量多、分布广,全面了解其遗传背景对于该地区猕猴资源的保护与合理利用具有重要的意义。本研究对来自四川8 个地理种群的231个不同猕猴个体的线粒体DNA 控制区部分序列进行了测定和群体分析,发现了110 个变异位点(22. 49% ),定义了56 种单倍型,其单倍型多样性(h)平均值为0. 686、核苷酸多样性(π)平均值为0. 01483,种群总体遗传多样性较高;进一步分析表明,8 个地理种群间存在着明显的遗传分化(F_st = 0. 70412,P < 0. 05),种群间基因交流较低(N_m < 1);系统发育树显示,四川猕猴8 个地理种群的单倍型基本上成簇分布在系统树上,与地理位置呈现一定的对应关系,说明四川猕猴具有明显的系统地理分布格局。地理隔离和人类活动可能是促使四川猕猴种群分化的主要因素。     

猕猴MHC-DPB1基因外显子2多态性研究

猕猴(Macaca mulatta)是最理想的医学实验灵长类动物, 且为国家二级保护动物。为了解中国猕猴主要组织相容复合体(Major histocompatibility complex, MHC)基因的遗传多态性背景, 为它们在生物医学研究中的应用及其遗传资源的保护提供一定的科学依据, 文章采用变性梯度凝胶电泳(Denaturing gradient gel electrophoresis, DGGE)和克隆测序技术分析了106个四川野生猕猴MHC-DPB1基因的exon 2, 共检测到21个Mamu-DPB1等位基因, 其中有15个为本研究中首次发现的新等位基因; 从整个大的猕猴群体(106个个体)来看, 等位基因频率最高的是Mamu-DPB1*30(0.1120); 单独从不同地理群体来看, 最高等位基因频率分别为: 小金-DPB1*30 (0.1120), 黑水-DPB1*04 (0.1702), 巴中-DPB1*32 (0.1613), 汉源-DPB1*30(0.1120), 九龙-DPB1*04(0.1139); 氨基酸序列比对发现, 猕猴Mamu-DPB1等位基因编码的氨基酸序列中, 有12个氨基酸残基变异位点表现出物种特异性, 其中有9个位于新发现的15个Mamu-DPB1等位基因氨基酸序列中; 不同物种来源的DPB1等位基因系统发生树表明, 猕猴与其近缘物种食蟹猴(Macaca fascicularis)的DPB1等位基因间存在着跨种多态(Trans-species polymorphism)现象。研究还表明, MHC-DPB1等位基因在中国猕猴群体和先前为主要研究对象的印度猕猴群体间具有较大的差异。     

Phylogenetic analysis of chinese rhesus macaques (Macaca mulatta) based on mitochondrial control region sequences

Between one and six subspecies of Chinese rhesus macaques (Macaca mulatta) have been proposed based on morphological differences and/or their geographic distribution. In this study, a 489 base pair fragment of the mitochondrial control region was amplified from 230 DNA samples collected from rhesus macaques in the Sichuan province in Western China. The fragment was then sequenced and aligned with 208 sequences from wild rhesus macaques, sampled throughout the species' geographic range in China downloaded from GenBank. Phylogenetic analysis of the 182 unique sequences identified among these samples divided Chinese rhesus macaques into two western haplogroups (haplogroups A and B) and three older eastern haplogroups (haplogroups C, D, and E), whose differentiation probably occurred during the penultimate glacial event. During the warming after the penultimate glacial event, haplogroups A, B, and E rapidly expanded and a relatively young subhaplogroup of haplogroup E, E', limited to Southern China but shared with Vietnamese rhesus macaques, was reintroduced from Indochina during the last glacial event. One haplotype most closely related to subhaplogroup E' probably represents the isolation of Hainan Island, to where it is restricted, from the mainland by the formation of the Qiongzhou Strait approximately 8,500 years ago. The distribution of haplogroups both informs the phylogeographic history of dispersal of Chinese rhesus macaques and has implications for their suitability as animal models in biomedical research.