共查询到20条相似文献，搜索用时 31 毫秒
1.
Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase 总被引：2172，自引：0，他引：2172
R K Saiki D H Gelfand S Stoffel S J Scharf R Higuchi G T Horn K B Mullis H A Erlich 《Science (New York, N.Y.)》1988,239(4839):487-491
A thermostable DNA polymerase was used in an in vitro DNA amplification procedure, the polymerase chain reaction. The enzyme, isolated from Thermus aquaticus, greatly simplifies the procedure and, by enabling the amplification reaction to be performed at higher temperatures, significantly improves the specificity, yield, sensitivity, and length of products that can be amplified. Single-copy genomic sequences were amplified by a factor of more than 10 million with very high specificity, and DNA segments up to 2000 base pairs were readily amplified. In addition, the method was used to amplify and detect a target DNA molecule present only once in a sample of 10(5) cells. 相似文献
2.
Large-scale and automated DNA sequence determination 总被引：28，自引：0，他引：28
DNA sequence analysis is a multistage process that includes the preparation of DNA, its fragmentation and base analysis, and the interpretation of the resulting sequence information. New technological advances have led to the automation of certain steps in this process and have raised the possibility of large-scale DNA sequencing efforts in the near future [for example, 1 million base pairs (Mb) per year]. New sequencing methodologies, fully automated instrumentation, and improvements in sequencing-related computational resources may render genome-size sequencing projects (100 Mb or larger) feasible during the next 5 to 10 years. 相似文献
3.
Kirkness EF Bafna V Halpern AL Levy S Remington K Rusch DB Delcher AL Pop M Wang W Fraser CM Venter JC 《Science (New York, N.Y.)》2003,301(5641):1898-1903
A survey of the dog genome sequence (6.22 million sequence reads; 1.5x coverage) demonstrates the power of sample sequencing for comparative analysis of mammalian genomes and the generation of species-specific resources. More than 650 million base pairs (>25%) of dog sequence align uniquely to the human genome, including fragments of putative orthologs for 18,473 of 24,567 annotated human genes. Mutation rates, conserved synteny, repeat content, and phylogeny can be compared among human, mouse, and dog. A variety of polymorphic elements are identified that will be valuable for mapping the genetic basis of diseases and traits in the dog. 相似文献
4.
Johnson WE Eizirik E Pecon-Slattery J Murphy WJ Antunes A Teeling E O'Brien SJ 《Science (New York, N.Y.)》2006,311(5757):73-77
Modern felid species descend from relatively recent (<11 million years ago) divergence and speciation events that produced successful predatory carnivores worldwide but that have confounded taxonomic classifications. A highly resolved molecular phylogeny with divergence dates for all living cat species, derived from autosomal, X-linked, Y-linked, and mitochondrial gene segments (22,789 base pairs) and 16 fossil calibrations define eight principal lineages produced through at least 10 intercontinental migrations facilitated by sea-level fluctuations. A ghost lineage analysis indicates that available felid fossils underestimate (i.e., unrepresented basal branch length) first occurrence by an average of 76%, revealing a low representation of felid lineages in paleontological remains. The phylogenetic performance of distinct gene classes showed that Y-chromosome segments are appreciably more informative than mitochondrial DNA, X-linked, or autosomal genes in resolving the rapid Felidae species radiation. 相似文献
5.
Non-Watson-Crick G.C and A.T base pairs in a DNA-antibiotic complex 总被引：11，自引：0，他引：11
G J Quigley G Ughetto G A van der Marel J H van Boom A H Wang A Rich 《Science (New York, N.Y.)》1986,232(4755):1255-1258
The structure of a DNA octamer d(GCGTACGC) cocrystallized with the bisintercalator antibiotic triostin A has been solved. The DNA forms an unwound right-handed double helix. Four base pairs are of the Watson-Crick type while four are Hoogsteen base pairs, including two A.T and two G.C base pairs. This is the first observation in an oligonucleotide of Hoogsteen G.C base pairs where the cystosine is protonated. It is likely that these also occur in solutions of DNA complexed to this antibiotic. 相似文献
6.
The genome sequence of the malaria mosquito Anopheles gambiae 总被引：2，自引：0，他引：2
Holt RA Subramanian GM Halpern A Sutton GG Charlab R Nusskern DR Wincker P Clark AG Ribeiro JM Wides R Salzberg SL Loftus B Yandell M Majoros WH Rusch DB Lai Z Kraft CL Abril JF Anthouard V Arensburger P Atkinson PW Baden H de Berardinis V Baldwin D Benes V Biedler J Blass C Bolanos R Boscus D Barnstead M Cai S Center A Chaturverdi K Christophides GK Chrystal MA Clamp M Cravchik A Curwen V Dana A Delcher A Dew I Evans CA Flanigan M Grundschober-Freimoser A Friedli L Gu Z Guan P Guigo R 《Science (New York, N.Y.)》2002,298(5591):129-149
7.
Metagenomic analysis of the human distal gut microbiome 总被引：2，自引：0，他引：2
Gill SR Pop M Deboy RT Eckburg PB Turnbaugh PJ Samuel BS Gordon JI Relman DA Fraser-Liggett CM Nelson KE 《Science (New York, N.Y.)》2006,312(5778):1355-1359
The human intestinal microbiota is composed of 10(13) to 10(14) microorganisms whose collective genome ("microbiome") contains at least 100 times as many genes as our own genome. We analyzed approximately 78 million base pairs of unique DNA sequence and 2062 polymerase chain reaction-amplified 16S ribosomal DNA sequences obtained from the fecal DNAs of two healthy adults. Using metabolic function analyses of identified genes, we compared our human genome with the average content of previously sequenced microbial genomes. Our microbiome has significantly enriched metabolism of glycans, amino acids, and xenobiotics; methanogenesis; and 2-methyl-d-erythritol 4-phosphate pathway-mediated biosynthesis of vitamins and isoprenoids. Thus, humans are superorganisms whose metabolism represents an amalgamation of microbial and human attributes. 相似文献
8.
Recognition of thymine adenine.base pairs by guanine in a pyrimidine triple helix motif 总被引：25，自引：0，他引：25
Oligonucleotide recognition offers a powerful chemical approach for the sequence-specific binding of double-helical DNA. In the pyrimidine-Hoogsteen model, a binding size of greater than 15 homopurine base pairs affords greater than 30 discrete sequence-specific hydrogen bonds to duplex DNA. Because pyrimidine oligonucleotides limit triple helix formation to homopurine tracts, it is desirable to determine whether oligonucleotides can be used to bind all four base pairs of DNA. A general solution would allow targeting of oligonucleotides (or their analogs) to any given sequence in the human genome. A study of 20 base triplets reveals that the triple helix can be extended from homopurine to mixed sequences. Guanine contained within a pyrimidine oligonucleotide specifically recognizes thymine.adenine base pairs in duplex DNA. Such specificity allows binding at mixed sites in DNA from simian virus 40 and human immunodeficiency virus. 相似文献
9.
A theoretical analysis of the reptational motion of DNA in a gel that includes the effects of molecular fluctuations has been used to explain the main features found in experiments involving periodic inversion of the electric field. The resonance-like decrease of the electrophoretic mobility as a function of pulse duration is related to transient "undershoots" in the orientation of the molecule, in agreement with recent experimental data. These features arise from a delicate interplay of internal and center of mass motion of the molecules under pulsed field conditions, and are important for the separation of DNA molecules in the size range 0.2 to 10 million base pairs. 相似文献
10.
以可乐猪、萝卜猪和香猪3个贵州地方猪种和欧洲猪种为材料,采用PCR-SSCP技术对脂蛋白脂酶（Lip-oprotein lipase,LPL）基因第8内含子的多态性进行了研究。以特异性PCR从样品基因组中扩增出163bp的DNA片段,经非变性聚丙烯酰胺凝胶电泳,从两个猪群体的LPL基因第8内含子中都检测到了AA基因型和AB基因型,未检测到BB基因型;欧洲猪品种和贵州地方猪品种均以AB基因型占优势;与欧洲猪品种相比,贵州地方猪品种的B等位基因频率明显较高,A等位基因频率则较低。 相似文献
11.
12.
L C Tsui M Buchwald D Barker J C Braman R Knowlton J W Schumm H Eiberg J Mohr D Kennedy N Plavsic 《Science (New York, N.Y.)》1985,230(4729):1054-1057
A polymorphic DNA marker has been found genetically linked, in a set of 39 human families, to an autosomal recessive gene that causes cystic fibrosis (CF), a disease affecting one in 2000 Caucasian children. The DNA marker (called D0CRI-917) is also linked to the PON locus, which by independent evidence is linked to the CF locus. The best estimates of the genetic distances are 5 centimorgans between the DNA marker and PON and 15 centimorgans between the DNA marker and the CF locus, meaning that the location of the disease gene has been narrowed to about 1 percent of the human genome (about 30 million base pairs). Although the data are consistent with the interpretation that a single locus causes cystic fibrosis, the possibility of genetic heterogeneity remains. The discovery of a linked DNA polymorphism is the first step in molecular analysis of the CF gene and its causative role in the disease. 相似文献
13.
White O Eisen JA Heidelberg JF Hickey EK Peterson JD Dodson RJ Haft DH Gwinn ML Nelson WC Richardson DL Moffat KS Qin H Jiang L Pamphile W Crosby M Shen M Vamathevan JJ Lam P McDonald L Utterback T Zalewski C Makarova KS Aravind L Daly MJ Minton KW Fleischmann RD Ketchum KA Nelson KE Salzberg S Smith HO Venter JC Fraser CM 《Science (New York, N.Y.)》1999,286(5444):1571-1577
The complete genome sequence of the radiation-resistant bacterium Deinococcus radiodurans R1 is composed of two chromosomes (2,648,638 and 412,348 base pairs), a megaplasmid (177,466 base pairs), and a small plasmid (45,704 base pairs), yielding a total genome of 3,284, 156 base pairs. Multiple components distributed on the chromosomes and megaplasmid that contribute to the ability of D. radiodurans to survive under conditions of starvation, oxidative stress, and high amounts of DNA damage were identified. Deinococcus radiodurans represents an organism in which all systems for DNA repair, DNA damage export, desiccation and starvation recovery, and genetic redundancy are present in one cell. 相似文献
14.
We describe a new molecular class of genetic-pairing system that has a native DNA backbone but has all four base pairs replaced by new, larger pairs. The base pairs include size-expanded analogs of thymine and of adenine, both extended by the width of a benzene ring (2.4 A). The expanded-diameter double helices are more thermodynamically stable than the Watson-Crick helix, likely because of enhanced base stacking. Structural data confirm a right-handed, double-stranded, and base-paired helical form. Because of the larger base size, all the pairs of this helical system are fluorescent, which suggests practical applications in detection of natural DNA and RNA. Our findings establish that there is no apparent structural or thermodynamic prohibition against genetic systems having sizes different from the natural one. 相似文献
15.
Echinomycin binding sites on DNA 总被引：25，自引：0，他引：25
The preferred binding sites of echinomycin on DNA can be determined by a method called "footprinting." A 32P end-labeled restriction fragment from pBR322 DNA is protected by binding to echinomycin, and cleaved by a synthetic DNA cleaving reagent, methidiumpropyl--EDTA . Fe(II); the DNA cleavage products are then subjected to high-resolution gel analyses. This method reveals that echinomycin has a binding site size of four base pairs. The strong binding sites for echinomycin contain the central two-base-pair sequence 5'-CG-3'. From an analysis of 15 echinomycin sites on 210 base pairs of DNA, key recognition elements for echinomycin are contained in the sequences (5'-3') ACGT and TCGT (A, adenine; C, cytosine; G, guanine; T, thymine). 相似文献
16.
为了对CbNPV ubiquitin基因序列进行分析,并对目前已知全基因组序列的杆状病毒的泛素氨基酸序列进行比较。从豆天蛾幼虫尸体中分离纯化豆天蛾核型多角体病毒(CbNPV),提取基因组DNA,进行基因组测序。序列分析发现一个长度为252 bp的读码框序列与泛素基因同源性很高,该基因编码83个氨基酸。氨基酸序列同源性分析结果表明,CbNPV泛素的氨基酸序列与20种杆状病毒泛素的氨基酸序列的同源性在66.3%~82.1%,并且维持泛素功能所必需的氨基酸序列,在大部分杆状病毒中也都保守存在。 相似文献
17.
Chromosomal region of the cystic fibrosis gene in yeast artificial chromosomes: a model for human genome mapping 总被引：33，自引：0，他引：33
A general strategy for cloning and mapping large regions of human DNA with yeast artificial chromosomes (YAC's) is described. It relies on the use of the polymerase chain reaction to detect DNA landmarks called sequence-tagged sites (STS's) within YAC clones. The method was applied to the region of human chromosome 7 containing the cystic fibrosis (CF) gene. Thirty YAC clones from this region were analyzed, and a contig map that spans more than 1,500,000 base pairs was assembled. Individual YAC's as large as 790 kilobase pairs and containing the entire CF gene were constructed in vivo by meiotic recombination in yeast between pairs of overlapping YAC's. 相似文献
18.
Trans-activator gene of human T-lymphotropic virus type III (HTLV-III) 总被引：204，自引：0，他引：204
19.
Stokstad E 《Science (New York, N.Y.)》2000,289(5479):530-531
Some 110 scientists from a range of disciplines gathered in the overcast British midlands for the 5th International Ancient DNA Conference, held here from 12 to 14 July. Among the attractions were new insights into the diets of early Americans gleaned from ancient human coprolites and intriguing reports of nuclear DNA and ancient viral sequences extracted from mammoth bones. 相似文献
20.
Site-specific oligonucleotide binding represses transcription of the human c-myc gene in vitro 总被引：80，自引：0，他引：80
M Cooney G Czernuszewicz E H Postel S J Flint M E Hogan 《Science (New York, N.Y.)》1988,241(4864):456-459