共查询到20条相似文献，搜索用时 31 毫秒
1.
Genomic and genetic definition of a functional human centromere 总被引：1，自引：0，他引：1
Schueler MG Higgins AW Rudd MK Gustashaw K Willard HF 《Science (New York, N.Y.)》2001,294(5540):109-115
The definition of centromeres of human chromosomes requires a complete genomic understanding of these regions. Toward this end, we report integration of physical mapping, genetic, and functional approaches, together with sequencing of selected regions, to define the centromere of the human X chromosome and to explore the evolution of sequences responsible for chromosome segregation. The transitional region between expressed sequences on the short arm of the X and the chromosome-specific alpha satellite array DXZ1 spans about 450 kilobases and is satellite-rich. At the junction between this satellite region and canonical DXZ1 repeats, diverged repeat units provide direct evidence of unequal crossover as the homogenizing force of these arrays. Results from deletion analysis of mitotically stable chromosome rearrangements and from a human artificial chromosome assay demonstrate that DXZ1 DNA is sufficient for centromere function. Evolutionary studies indicate that, while alpha satellite DNA present throughout the pericentromeric region of the X chromosome appears to be a descendant of an ancestral primate centromere, the current functional centromere based on DXZ1 sequences is the product of the much more recent concerted evolution of this satellite DNA. 相似文献
2.
Isolation and mapping of a mouse complementary DNA sequence (mouse Y-finger) encoding a multiple, potential zinc-binding, finger protein homologous to the candidate human testis-determining factor gene is reported. Four similar sequences were identified in Hind III-digested mouse genomic DNA. Two (7.2 and 2.0 kb) were mapped to the Y chromosome. Only the 2.0-kb fragment, however, was correlated with testis determination. Polymerase chain reaction analysis suggests both Y loci are transcribed in adult testes. A 3.6-kb fragment was mapped to the X chromosome between the T16H and T6R1 translocation breakpoints, and a fourth (6.0 kb) was mapped to chromosome 10. Hence, mYfin sequences have been duplicated several times in the mouse, although they are not duplicated in humans. 相似文献
3.
Cloning of human androgen receptor complementary DNA and localization to the X chromosome 总被引：55，自引：0，他引：55
D B Lubahn D R Joseph P M Sullivan H F Willard F S French E M Wilson 《Science (New York, N.Y.)》1988,240(4850):327-330
The androgen receptor (AR) mediates the actions of male sex steroids. Human AR genomic DNA was cloned from a flow-sorted human X chromosome library by using a consensus nucleotide sequence from the DNA-binding domain of the family of nuclear receptors. The AR gene was localized on the human X chromosome between the centromere and q13. Cloned complementary DNA, selected with an AR-specific oligonucleotide probe, was expressed in monkey kidney (COS) cells and yielded a high-affinity androgen-binding protein with steroid-binding specificity corresponding to that of native AR. A predominant messenger RNA species of 9.6 kilobases was identified in human, rat, and mouse tissues known to contain AR and was undetectable in tissues lacking AR androgen-binding activity, including kidney and liver from androgen-insensitive mice. The deduced amino acid sequence of AR within the DNA-binding domain has highest sequence identity with the progesterone receptor. 相似文献
4.
A human Y-linked DNA polymorphism and its potential for estimating genetic and evolutionary distance 总被引：29，自引：0，他引：29
M Casanova P Leroy C Boucekkine J Weissenbach C Bishop M Fellous M Purrello G Fiori M Siniscalco 《Science (New York, N.Y.)》1985,230(4732):1403-1406
A human DNA sequence (p12f2), derived from a partial Y-chromosome genomic library and showing homology with the X and Y chromosomes and with an undetermined number of autosomes, detected two Y-specific restriction fragment length variants on male DNA that had been digested with Taq I and Eco RI. These variants may have been generated through a deletion-insertion mechanism and their pattern of holoandric transmission indicates that they represent a two-allele Y-linked polymorphism (RFLP). By means of DNA from patients with inborn deletions in chromosome Y, this polymorphic DNA site was mapped to the interval Yq11.1-Yq11.22. The frequency of the rarest allele was about 35 percent in Algerian and Sardinian human males, whereas it was only 4 percent among Northern Europeans. The p12f2 probe also detected Y-specific DNA fragments in the gorilla and chimpanzee. In view of the monosomy of the Y chromosome in mammalian species, Y-linked RFLP's may prove to be more useful than autosomal or X-linked markers in estimating genetic distances within and between species. 相似文献
5.
应用PCR方法通过毛发进行哺乳动物性别鉴定 总被引：15，自引：2，他引：13
以哺乳动物的新鲜毛发和在冷冻条件下保存４个月的毛发作为材料。从毛囊中提取ＤＮＡ,应用ＰＣＲ方法扩增ＳＲＹ基因片段,ＺＦＸ／ＺＦＹ基因片段,探讨哺乳动物性别鉴定的最优化ＰＣＲ方案。结果表明,毛发ＤＮＡ和血液ＤＮＡ均可获得相同的扩增结果。证实了毛发作为ＰＣＲ扩增材料是可靠的。对不同材料量、不同模板量、不同提取方法进行比较,优化实验方案,达到１根毛发用Ｃｈｅｌｅｘ－１００提取得到的ＤＮＡ即可以成功地进行 相似文献
6.
Genomic amplification with transcript sequencing 总被引：48，自引：0，他引：48
7.
Interferon-beta-related DNA is dispersed in the human genome 总被引：6，自引：0，他引：6
A D Sagar P B Sehgal L T May M Inouye D L Slate L Shulman F H Ruddle 《Science (New York, N.Y.)》1984,223(4642):1312-1315
Interferon-beta 1 (IFN-beta 1) complementary DNA was used as a hybridization probe to isolate human genomic DNA clones lambda B3 and lambda B4 from a human genomic DNA library. Blot-hybridization procedures and partial nucleotide sequencing revealed that lambda B3 is related to IFN-beta 1 (and more distantly to IFN-alpha 1). Analyses of DNA obtained from a panel of human-rodent somatic cell hybrids that were probed with DNA derived from lambda B3 showed that lambda B3 is on human chromosome 2. Similar experiments indicated that lambda B4 is not on human chromosomes 2, 5, or 9. The finding that DNA related to the IFN-beta 1 gene (and IFN-alpha 1 gene) is dispersed in the human genome raises new questions about the origins of the interferon genes. 相似文献
8.
Complementary DNA sequencing: expressed sequence tags and human genome project 总被引：227，自引：0，他引：227
M D Adams J M Kelley J D Gocayne M Dubnick M H Polymeropoulos H Xiao C R Merril A Wu B Olde R F Moreno 《Science (New York, N.Y.)》1991,252(5013):1651-1656
Automated partial DNA sequencing was conducted on more than 600 randomly selected human brain complementary DNA (cDNA) clones to generate expressed sequence tags (ESTs). ESTs have applications in the discovery of new human genes, mapping of the human genome, and identification of coding regions in genomic sequences. Of the sequences generated, 337 represent new genes, including 48 with significant similarity to genes from other organisms, such as a yeast RNA polymerase II subunit; Drosophila kinesin, Notch, and Enhancer of split; and a murine tyrosine kinase receptor. Forty-six ESTs were mapped to chromosomes after amplification by the polymerase chain reaction. This fast approach to cDNA characterization will facilitate the tagging of most human genes in a few years at a fraction of the cost of complete genomic sequencing, provide new genetic markers, and serve as a resource in diverse biological research fields. 相似文献
9.
Molecular genetics of human color vision: the genes encoding blue, green, and red pigments 总被引：86，自引：0，他引：86
Human color vision is based on three light-sensitive pigments. The isolation and sequencing of genomic and complementary DNA clones that encode the apoproteins of these three pigments are described. The deduced amino acid sequences show 41 +/- 1 percent identity with rhodopsin. The red and green pigments show 96 percent mutual identity but only 43 percent identity with the blue pigment. Green pigment genes vary in number among color-normal individuals and, together with a single red pigment gene, are proposed to reside in a head-to-tail tandem array within the X chromosome. 相似文献
10.
参考家猪(X89007)MyoG基因序列设计2对特异引物,用PCR方法首次从野猪基因组中扩增出2个大小分别约为1.6 kb和1.0 kb的DNA片段,其PCR产物经pMD-18T载体转化感受态DH5α株大肠杆菌,获得重组克隆子。DNA测序和序列拼接表明:野猪肌细胞生成素基因DNA序列长2 466bp,含完整的3个外显子和2个内含子,与家猪、牛、马、狗、小鼠和人的MyoG基因的cDNA序列同源性分别为99.8%、92.4%、92.7%、89.7%、90.8%和94%,其cDNA编码氨基酸序列与家猪、牛、马、狗、小鼠和人的同源性分别为100%、96.4%、95.9%、94.1%、96.4%、96.8%;对野猪MyoG基因组序列与9个家猪品种的相应同源序列进行比较,检出16个核苷酸变异位点,且其中有5个为家猪中不具有的新变异位点,其变异位点主要发生在内含子部分,尤其是内含子1中的变异位点比例最大(11个)。这些结果表明,野猪肌细胞生成素基因的编码序列在进化过程中是高度保守的,而内含子部分尤其是第1内含子具有丰富的序列多态性。对117个限制性酶切位点扫描分析发现,野猪MyoG基因核苷酸序列中含有78个酶切位点,其中有5个酶切位点包含变异位点,尤其是1 153位点的T突变成G所产生的SmaI(CCC/GGG)或XmaI(C/CCGGG)酶切位点为野猪所特有。所测DNA序列已提交到GenBank中,获得的序列号为:FJ356697。 相似文献
11.
The fragile X site in somatic cell hybrids: an approach for molecular cloning of fragile sites 总被引：17，自引：0，他引：17
Fragile X syndrome is a common form of mental retardation associated with a fragile site on the human X chromosome. Although fragility at this site is usually evident as a nonstaining chromatid gap, it remains unclear whether or not actual chromosomal breakage occurs. By means of somatic cell hybrids containing either a normal human X or a fragile X chromosome and utilizing two genes that flank the fragile site as markers of chromosome integrity, segregation of these markers was shown to be more frequent if they encompass the fragile site under appropriate culture conditions. Hybrid cells that reveal marker segregation were found to contain rearranged X chromosomes involving the region at or near the fragile site, thus demonstrating true chromosomal breakage within this area. Two independent translocation chromosomes were identified involving a rodent chromosome joined to the human X at the location of the fragile site. DNA analysis of closely linked, flanking loci was consistent with the position of the breakpoint being at or very near the fragile X site. Fragility at the translocation junctions was observed in both hybrids, but at significantly lower frequencies than that seen in the intact X of the parental hybrid. This observation suggests that the human portion of the junctional DNA may contain part of a repeated fragility sequence. Since the translocation junctions join heterologous DNA, the molecular cloning of the fragile X sequence should now be possible. 相似文献
12.
The sequence of the human genome 总被引：8，自引：0，他引：8
Venter JC Adams MD Myers EW Li PW Mural RJ Sutton GG Smith HO Yandell M Evans CA Holt RA Gocayne JD Amanatides P Ballew RM Huson DH Wortman JR Zhang Q Kodira CD Zheng XH Chen L Skupski M Subramanian G Thomas PD Zhang J Gabor Miklos GL Nelson C Broder S Clark AG Nadeau J McKusick VA Zinder N Levine AJ Roberts RJ Simon M Slayman C Hunkapiller M Bolanos R Delcher A Dew I Fasulo D Flanigan M Florea L Halpern A Hannenhalli S Kravitz S Levy S Mobarry C Reinert K Remington K Abu-Threideh J Beasley E 《Science (New York, N.Y.)》2001,291(5507):1304-1351
13.
Clustering of human H1 and core histone genes 总被引：9，自引：0，他引：9
N Carozzi F Marashi M Plumb S Zimmerman A Zimmerman L S Coles J R Wells G Stein J Stein 《Science (New York, N.Y.)》1984,224(4653):1115-1117
An H1 histone gene was isolated from a 15-kilobase human DNA genomic sequence. The presence of H2A, H2B, H3, and H4 genes in this same 15-kilobase fragment indicates that mammalian core and H1 histone genes are clustered. 相似文献
14.
利用y-型高分子量谷蛋白亚基的特异引物,对阿拉拉特小麦(PI427305)的基因组DNA进行PCR扩增,得到大小为2.2 kb的目的条带,将该条带回收纯化并克隆到pMD18-T载体中,经梯度亚克隆测序拼接,得到编码区的全序列为2 202 bp(GenBank登录号:HM131806),共编码732个氨基酸。它与1Gy7*序列的同源性高达99%,而且氨基酸序列结构与大多数y-型亚基相同,推断该基因为1Gy。1Gy的分子量比1Gy7*稍小,迁移率比1Gy7*慢。与小麦属其他基因组编码的y-型亚基相比,其在靠近C端的重复区多了一个半胱氨酸残基。利用在线PSIPRED对其二级结构预测结果显示,其重复区主要是无规则卷曲结构。这些结构都可能使得1Gy对小麦加工品质产生正面影响。 相似文献
15.
Construction of a general human chromosome jumping library, with application to cystic fibrosis 总被引：22，自引：0，他引：22
F S Collins M L Drumm J L Cole W K Lockwood G F Vande Woude M C Iannuzzi 《Science (New York, N.Y.)》1987,235(4792):1046-1049
In many genetic disorders, the responsible gene and its protein product are unknown. The technique known as "reverse genetics," in which chromosomal map positions and genetically linked DNA markers are used to identify and clone such genes, is complicated by the fact that the molecular distances from the closest DNA markers to the gene itself are often too large to traverse by standard cloning techniques. To address this situation, a general human chromosome jumping library was constructed that allows the cloning of DNA sequences approximately 100 kilobases away from any starting point in genomic DNA. As an illustration of its usefulness, this library was searched for a jumping clone, starting at the met oncogene, which is a marker tightly linked to the cystic fibrosis gene that is located on human chromosome 7. Mapping of the new genomic fragment by pulsed field gel electrophoresis confirmed that it resides on chromosome 7 within 240 kilobases downstream of the met gene. The use of chromosome jumping should now be applicable to any genetic locus for which a closely linked DNA marker is available. 相似文献
16.
Direct cloning and sequence analysis of enzymatically amplified genomic sequences 总被引：120，自引：0，他引：120
A method is described for directly cloning enzymatically amplified segments of genomic DNA into an M13 vector for sequence analysis. A 110-base pair fragment of the human beta-globin gene and a 242-base pair fragment of the human leukocyte antigen DQ alpha locus were amplified by the polymerase chain reaction method, a procedure based on repeated cycles of denaturation, primer annealing, and extension by DNA polymerase I. Oligonucleotide primers with restriction endonuclease sites added to their 5' ends were used to facilitate the cloning of the amplified DNA. The analysis of cloned products allowed the quantitative evaluation of the amplification method's specificity and fidelity. Given the low frequency of sequence errors observed, this approach promises to be a rapid method for obtaining reliable genomic sequences from nanogram amounts of DNA. 相似文献
17.
Characterization and molecular cloning of a human parvovirus genome 总被引：25，自引：0，他引：25
The genome of the small human virus serologically associated with erythrocyte aplasia and erythema infectiosum (fifth disease) is shown to be a linear, nonpermuted, single-stranded DNA molecule with self-priming hairpin termini, properties which are characteristic of the genomes of the family Parvoviridae. This human parvovirus chromosome was molecularly cloned into bacterial plasmid vectors and the cloned DNA was used to explore its relatedness to other mammalian parvovirus serotypes by DNA:DNA hybridization. It is not related to the human adeno-associated viruses but does show a distant evolutionary relationship to genomes of the helper-independent parvoviruses of rodents. This strongly suggests that it is an autonomous parvovirus, and as such is the first example of a member of this group of common animal pathogens to cause disease in man. 相似文献
18.
19.
Identification of a chromosome 18q gene that is altered in colorectal cancers 总被引：141，自引：0，他引：141
E R Fearon K R Cho J M Nigro S E Kern J W Simons J M Ruppert S R Hamilton A C Preisinger G Thomas K W Kinzler 《Science (New York, N.Y.)》1990,247(4938):49-56
Allelic deletions involving chromosome 18q occur in more than 70 percent of colorectal cancers. Such deletions are thought to signal the existence of a tumor suppressor gene in the affected region, but until now a candidate suppressor gene on this chromosomal arm had not been identified. A contiguous stretch of DNA comprising 370 kilobase pairs (kb) has now been cloned from a region of chromosome 18q suspected to reside near this gene. Potential exons in the 370-kb region were defined by human-rodent sequence identities, and the expression of potential exons was assessed by an "exon-connection" strategy based on the polymerase chain reaction. Expressed exons were used as probes for cDNA screening to obtain clones that encoded a portion of a gene termed DCC; this cDNA was encoded by at least eight exons within the 370-kb genomic region. The predicted amino acid sequence of the cDNA specified a protein with sequence similarity to neural cell adhesion molecules and other related cell surface glycoproteins. While the DCC gene was expressed in most normal tissues, including colonic mucosa, its expression was greatly reduced or absent in most colorectal carcinomas tested. Somatic mutations within the DCC gene observed in colorectal cancers included a homozygous deletion of the 5' end of the gene, a point mutation within one of the introns, and ten examples of DNA insertions within a 0.17-kb fragment immediately downstream of one of the exons. The DCC gene may play a role in the pathogenesis of human colorectal neoplasia, perhaps through alteration of the normal cell-cell interactions controlling growth. 相似文献
20.
Benos PV Gatt MK Ashburner M Murphy L Harris D Barrell B Ferraz C Vidal S Brun C Demailles J Cadieu E Dreano S Gloux S Lelaure V Mottier S Galibert F Borkova D Minana B Kafatos FC Louis C Sidén-Kiamos I Bolshakov S Papagiannakis G Spanos L Cox S Madueño E de Pablos B Modolell J Peter A Schöttler P Werner M Mourkioti F Beinert N Dowe G Schäfer U Jäckle H Bucheton A Callister DM Campbell LA Darlamitsou A Henderson NS McMillan PJ Salles C Tait EA Valenti P Saunder RD Glover DM 《Science (New York, N.Y.)》2000,287(5461):2220-2222
One of the rewards of having a Drosophila melanogaster whole-genome sequence will be the potential to understand the molecular bases for structural features of chromosomes that have been a long-standing puzzle. Analysis of 2.6 megabases of sequence from the tip of the X chromosome of Drosophila identifies 273 genes. Cloned DNAs from the characteristic bulbous structure at the tip of the X chromosome in the region of the broad complex display an unusual pattern of in situ hybridization. Sequence analysis revealed that this region comprises 154 kilobases of DNA flanked by 1.2-kilobases of inverted repeats, each composed of a 350-base pair satellite related element. Thus, some aspects of chromosome structure appear to be revealed directly within the DNA sequence itself. 相似文献