渤海黑牛和日本和牛mtDNAD-loop区遗传变异研究

研究渤海黑牛和日本和牛线粒体DNA控制区(mtDNA D-loop)的遗传变异。采用DNA测序技术测定了这2个品种17个个体mtDNA D-loop区全序列,从Genbank下载了31头日本和牛、9头渤海黑牛、2头欧洲普通牛、2头瘤牛以及21个其他中外黄牛品种mtDNA D-loop区全序列,用lasergene和Mega 4.0.1软件对所得的渤海黑牛和日本和牛57个个体的序列进行了遗传多样性研究,并对渤海黑牛、日本和牛、欧洲普通牛和瘤牛的mtDNAD-loop区全序列进行了同源性分析。以水牛为外群,对23个中外黄牛品种的聚类分析显示它们聚为2类:欧洲型原牛和瘤牛型原牛。渤海黑牛含有这2类的遗传背景,受到过瘤牛血缘渐渗的影响。而日本和牛则来源于欧洲原牛。结论:渤海黑牛和日本和牛mtDNA D-loop区具有丰富的遗传多样性,二者的亲缘关系很近;渤海黑牛含有普通牛和瘤牛的血统、受到过瘤牛血缘渐渗的影响;日本和牛属普通牛类型,欧洲原牛、朝鲜牛和我国北方的部分牛种对于日本和牛的形成可能都起到丰富种群的作用。     

基于Cytb基因的东印度洋鸢乌贼系统发育关系和种群遗传结构

通过测定东印度洋北部、赤道、南部3个鸢乌贼（Sthenoteuthis oualaniensis）群体的线粒体DNA细胞色素b（Cytb）基因序列,分析了东印度洋鸢乌贼的系统发育关系。研究表明,东印度洋鸢乌贼整体具有高单倍型多样性（h=0.939）和高核苷酸多样性（π=0.015）。3个鸢乌贼群体间的F_st值为0.661,群体间遗传分化水平很高,具有明显的遗传分化现象,可分为以东印度洋北部群体为主的谱系1（东印度洋北部谱系）和主要由东印度洋赤道与南部群体组成的谱系2（东印度洋南部谱系）,且谱系2经历过群体扩张。东印度洋巨大的南北水文差异可能是鸢乌贼产生南北谱系分化的原因,在东印度洋鸢乌贼资源的利用和管理方面应考虑南北谱系的差异和特点。     

用M13小卫星探针发现猪DNA指纹

用Ｍ１３ｍｐ１８含基因Ⅲ区的ＣｌａⅠ限制性酶切片段为探针，与猪基因组ＤＮＡ的ＨｉｎｆⅠ限制性酶切产物进行杂交，发现了ＤＮＡ指纹，平均图带数为１７．３１条。说明Ｍ１３ｍｐ１８小卫星序列可作为猪ＤＮＡ指纹图谱分析的探针。     

榆中县黄瓜根结线虫鉴定及根系定殖效果测定

为了明确甘肃榆中县温室大棚黄瓜根结线虫种类及根际定殖情况,采用形态和分子生物学鉴定其种类,并室内测定和显微观察其在根际定殖效果。结果表明,甘肃榆中县黄瓜根结线虫不同虫态形态、大小和雌虫会阴花纹特征等均具有典型南方根结线虫(Meloidogyne incognita)形态特征。基于线粒体COI基因序列比对和构建系统发育树,鉴定发现榆中县黄瓜根结线虫线粒体COI基因序列与已报道的南方根结线虫台湾株系(KU517167)聚为一类,其相似性为99.75%;根结线虫多序列特异位点鉴定发现其与南方根结线虫孟加拉国株系(MN196556)和南方根结线虫印度株系(MN728697)聚为一类,相似性分别为100%和96.91%。经致病性测定发现,待接种根结线虫2龄幼虫30 d后其能够侵染黄瓜根系并形成大量根结,经染色发现黄瓜幼苗根系组织内寄生大量线形蠕虫、雌虫和卵囊,且镜检观察发现其形态与田间采集后分离的线虫形态一致。综上可知,引起甘肃省榆中县黄瓜根结线虫病的病原为南方根结线虫。     

Comment on "Population size does not influence mitochondrial genetic diversity in animals"

Bazin et al. (Reports, 28 April, 2006, p. 570) found no relationship between mitochondrial DNA (mtDNA) diversity and population size when comparing across large groups of animals. We show empirically that species with smaller populations, as represented by eutherian mammals, exhibit a positive correlation between mtDNA and allozyme variation, suggesting that mtDNA diversity may correlate with population size in these animals.     

Reconstructing the origin of Andaman Islanders

The origin of the Andaman "Negrito" and Nicobar "Mongoloid" populations has been ambiguous. Our analyses of complete mitochondrial DNA sequences from Onges and Great Andaman populations revealed two deeply branching clades that share their most recent common ancestor in founder haplogroup M, with lineages spread among India, Africa, East Asia, New Guinea, and Australia. This distribution suggests that these two clades have likely survived in genetic isolation since the initial settlement of the islands during an out-of-Africa migration by anatomically modern humans. In contrast, Nicobarese sequences illustrate a close genetic relationship with populations from Southeast Asia.     

Restriction endonuclease analysis of mitochondrial DNA from normal and Texas cytoplasmic male-sterile maize

Mitochondrial DNA from normal and cytoplasmic male-sterile maize was digested with restriction endonucleases RI from Escherichia coli or dIII from Hemophilus influenzae. Electrophoresis of resulting fragments revealed distinctions between the two cytoplasmic types. These distinctions suggest that factors responsible or cytoplasmic male sterility are located in the mitochondrial DNA, and that the mitochondrial genome is not inherited paternally.     

一种含硒鬼臼衍生物通过引起线粒体膜电位下降诱导SMMC-7721细胞凋亡

[目的]为从植物中提取和开发新药提供依据。[方法]研究含硒鬼臼衍生物4′-去甲基-4-脱羟基-4-硒代-对甲苯基-β-表鬼臼毒素(SEP)对SMMC-7721细胞凋亡的诱导,并分析SMMC-7721细胞线粒体膜电位的变化。[结果]SEP可抑制SMMC-7721的增殖,且有明显的剂量效应。经SEP处理后,SMMC-7721细胞形态明显改变,出现了皱缩、变小等疑似凋亡的特征。M30抗体检测表明SMMC-7721细胞凋亡率随SEP浓度的增加而增加,证实SEP可诱导SMMC-7721细胞凋亡。低浓度SEP(10、20μmol/L)可使SMMC-7721细胞阻滞在G2/M期,而40μmol/LSEP则使阻滞在S期的细胞数明显升高,说明细胞DNA发生了损伤,这很可能引起线粒体凋亡途径的产生。不同浓度SEP处理均可使SMMC-7721细胞线粒体膜电位的下降。[结论]SMMC-7721细胞凋亡可能是通过线粒体途径产生的。     

鹿科动物线粒体DNA序列差异和系统进化关系

从Gen Bank数据库中获得20种鹿科动物的线粒体DNA序列全长,分析了序列碱基含量和遗传距离关系,并构建系统进化树,探讨了鹿科动物的系统进化关系。序列分析表明:鹿科动物线粒体DNA全长为16 305~16 482 bp,A+T含量约占62.58%,各物种间遗传距离为0.014~0.164。系统进化分析表明:驼鹿在鹿科动物中可能是最古老的;麋鹿与鹿属亲缘关系较近,支持将麋鹿划到鹿属的观点;泽鹿与花鹿属的斑鹿和豚鹿先聚到一起,支持将泽鹿划到花鹿属中;麂亚科中小麂可能是最原始的,赤麂和黑麂亲缘关系较近;獐与狍亲缘关系更近,建议将獐与狍划归到一个亚科。     

基于棉花BAC文库的线粒体DNA的提取方法

以棉花的黄化苗为材料，根据棉花自身的特点，利用差速离心的方法分离线粒体后再经低熔点琼脂糖包埋裂解提取了完整的mtDNA。其质量完全可以满足BAC文库构建中限制性酶切、PCR检测、分子杂交筛选等实验的要求。     

Nuclear and mitochondrial DNA comparisons reveal extreme rate variation in the molecular clock

The discovery that the rate of evolution of vertebrate mitochondrial DNA is rapid, compared to the rate for vertebrate nuclear DNA, has resulted in its widespread use in evolutionary studies. Comparison of mitochondrial and nuclear DNA divergences among echinoid and vertebrate taxa of similar ages indicates that the rapid rate of vertebrate mitochondrial DNA evolution is, in part, an artifact of a widely divergent rate of nuclear DNA evolution. This disparity in relative rates of mitochondrial and nuclear DNA divergence suggests that the controls and constraints under which the mitochondrial and nuclear genomes operate are evolving independently, and provides evidence that is independent of fossil dating for a robust rejection of a generalized molecular clock hypothesis of DNA evolution.     

URF6, last unidentified reading frame of human mtDNA, codes for an NADH dehydrogenase subunit

The polypeptide encoded in URF6, the last unassigned reading frame of human mitochondrial DNA, has been identified with antibodies to peptides predicted from the DNA sequence. Antibodies prepared against highly purified respiratory chain NADH dehydrogenase from beef heart or against the cytoplasmically synthesized 49-kilodalton iron-sulfur subunit isolated from this enzyme complex, when added to a deoxycholate or a Triton X-100 mitochondrial lysate of HeLa cells, specifically precipitated the URF6 product together with the six other URF products previously identified as subunits of NADH dehydrogenase. These results strongly point to the URF6 product as being another subunit of this enzyme complex. Thus, almost 60% of the protein coding capacity of mammalian mitochondrial DNA is utilized for the assembly of the first enzyme complex of the respiratory chain. The absence of such information in yeast mitochondrial DNA dramatizes the variability in gene content of different mitochondrial genomes.     

Conferring operator specificity on restriction endonucleases

Mapping and manipulation of very large genomes, including the human genome, would be facilitated by the availability of a DNA cleavage method with very high site specificity. Therefore, a general method was devised that extends the effective recognition sequences well beyond the present 8-base pair limit by combining the specificity of the restriction endonuclease with that of another sequence-specific protein that binds tightly to DNA. It was shown that the tightly binding lac or lambda repressor protects a restriction site within the operator from specific modification methylases, M.Hha I or M.Hph I, while all other similar sites are methylated and thus rendered uncleavable. A plasmid containing a symmetric lac operator was specifically cleaved by Hha I, only at the site within the operator, after M.Hha I methylation in the presence of the lac repressor, whereas the remaining 31 Hha I sites on this plasmid were methylated and thus not cleaved. Analogous results were obtained with the Hae II site within the lac operator, which was similarly protected by the lac repressor, and with the Hph I site within the phage lambda oL operator, which was protected by lambda repressor from M.Hph I methylation.     

Microsome-associated DNA

Deoxyribonucleic acid has been isolated from the microsomes of mouse liver homogenates under conditions designed to prevent or greatly reduce mitochondrial and nuclear contamination. The DNA rapidly incorporates tritiated thymidine, and this, together with its reannealing characteristics after thermal denaturation, shows that it is not mitochondrial or typically nuclear DNA.     

Linkage disequilibrium and recombination in hominid mitochondrial DNA

The assumption that human mitochondrial DNA is inherited from one parent only and therefore does not recombine is questionable. Linkage disequilibrium in human and chimpanzee mitochondrial DNA declines as a function of the distance between sites. This pattern can be attributed to one mechanism only: recombination.     

Mitochondrial DNA size variation within individual crickets

The mitochondrial DNA's of two closely related cricket species (genus Gryllus) share a size polymorphism as evidenced by analysis of restriction fragment patterns. Moreover, 12 of 100 field-collected crickets are heteroplasmic, that is these individuals have more than one size class of mitochondrial DNA. No heteroplasmy for restriction site variation is observed. Intraindividual variation in cricket mitochondrial DNA provides a useful marker for studying the transmission genetics of mitochondrial DNA. Available data on patterns of variation in mothers and offspring suggest that random segregation of mitochondrial DNA variants does not occur rapidly in cricket germ-cell lineages.     

Mitochondrial malate dehydrogenase: a new genetic polymorphism in man

Starch-gel electrophoresis patterns of malate dehydrogenase from human tissue indicate a new genetic polymorphism for the mitochondrial form of the enzyme. Studies of families showed simple Mendelian segregation rather than maternal inheritance, suggesting that not all mitochondrial proteins are coded by mitochondrial DNA.     

Method for Isolating Mitochondrial DNA from Etiolated Tissue of Cabbage

Isolation of high-quality mitochondrial DNA（mtDNA） is an important premise for researching molecular mechanisms in cytoplasmic male sterility of cabbage（Brassica oleracea L.var.capitata）. An efficient protocol for separation and purification of mitochondria and extraction of mitochondrial DNA（mtDNA） from etiolated tissues of cabbage was developed. We took a method combined mannitol density gradient with differential centrifugation, selected appropriate rotational speed, extended DNase I treating time and changed mitochondria cracking condition. The results showed that the extracted mitochondria in this protocol had complete structure, appeared to ellipsoid and had not been contaminated with other impurities under the Jannus Green B staining. The isolated mitochondrial DNA had high purity and yield through detecting the optical density, nuclear specific primer PCR and agarose gel electrophoresis. The results indicated that mitochondrial DNA extracted by this protocol had high quality and enabled to be used in futher genetic studies.     

中国飞蝗线粒体COI基因片段的扩增

对中国飞蝗各亚种线粒体DNA的COI基因片段进行了扩增并测序。结果表明,扩增产物为单一带,大小约为1.3kb。对此扩增产物序列分析证明所扩增的COI基因序列是一个含混不清的序列,说明在中国飞蝗不同亚种的基因组DNA中存在线粒体的假基因序列,因此以飞蝗基因组DNA为模板PCR扩出的mtDNA COI基因片段不适宜用作分析飞蝗种群遗传和系统发育的分子标记。     

Comment on "The great Sumatra-Andaman earthquake of 26 December 2004"

Lay et al. (Research Articles, 20 May 2005, p. 1127) estimated a 600-km length for the tsunami source region. Adding tide-gauge data from Paradip, the northernmost of the Indian east-coast stations and therefore the most critical constraint on the northern extent of the source, we estimate that its length was greater by approximately 30%.