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Binding of 5-bromouracil-containing S/MAR DNA to the nuclear matrix. 含5-溴酸的S/MAR DNA与核基质的结合。
H. Ogino, M. Fujii, W. Satou, Toshikazu Suzuki, E. Michishita, D. Ayusawa
Substitution of thymine with 5-bromouracil in DNA is known to change interaction between DNA and proteins, thereby inducing various biological phenomena. We hypothesize that A/T-rich scaffold/nuclear matrix attachment region (S/MAR) sequences are involved in the effects of 5-bromodeoxyuridine. We examined an interaction between DNA containing an intronic S/MAR sequence of the immunoglobulin heavy chain gene and nuclear halos prepared from HeLa cells. Upon substitution with 5-bromouracil, the S/MAR DNA bound more tightly to the nuclear halos. The multi-functional nuclear matrix protein YY1 was also found to bind more strongly to 5-bromouracil-substituted DNA containing its recognition motif. These results are consistent with the above hypothesis.
已知胸腺嘧啶在DNA中被5-溴酸取代会改变DNA与蛋白质之间的相互作用,从而诱发各种生物现象。我们推测富含A/ t的支架/核基质附着区(S/MAR)序列参与了5-溴脱氧尿苷的作用。我们检测了含有免疫球蛋白重链基因内含子S/MAR序列的DNA与HeLa细胞制备的核晕之间的相互作用。与5-溴醛酸取代后,S/MAR DNA与核晕结合更紧密。多功能核基质蛋白YY1也被发现与含有其识别基序的5-溴酰取代的DNA结合更强。这些结果与上述假设一致。
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引用次数: 20
Structural analysis of a Lotus japonicus genome. II. Sequence features and mapping of sixty-five TAC clones which cover the 6.5-mb regions of the genome. 日本莲基因组的结构分析。2覆盖基因组6.5 mb区域的65个TAC克隆的序列特征和图谱。
Yasukazu Nakamura, T. Kaneko, E. Asamizu, Tomohiko Kato, Shusei Sato, S. Tabata
Sixty-five TAC (transformation-competent artificial chromosomes) clones were selected from a genomic library of Lotus japonicus accession MG-20 based on the sequence information of expressed sequences tags (ESTs), cDNA and gene information, and their nucleotide sequences were determined. The average insert size of the TAC clone was approximately 100 kb, and the total length of the sequenced regions in this study is 6,556,100 bp. Together with the nucleotide sequences of 56 TAC clones previously reported, the regions sequenced so far total 12,029,295 bp. By comparison with the sequences in protein and EST databases and by analysis with computer programs for gene modeling, a total of 711 potential protein-encoding genes with known or predicted functions, 239 gene segments and 90 pseudogenes were identified in the newly sequenced regions. The average gene density assigned so far was 1 gene/9140 bp. The average length of the assigned genes was 2.6 kb, which is considerably larger than that assigned in the Arabidopsis thaliana genome (1.9 kb for 6451 genes). Introns were identified in approximately 73% of the potential genes, and the average number and length of the introns per gene were 3.4 and 377 bp, respectively. Simple sequence repeat length polymorphism (SSLP) or derived cleaved amplified polymorphic sequence (dCAPS) markers were generated based on the nucleotide sequences of the genomic clones obtained, and each clone was mapped onto the linkage map using the F2 mapping population derived from a cross of two accessions of L. japonicus, Gifu B-129 and Miyakojima MG-20. The sequence data, gene information and mapping information are available through the World Wide Web at http://www.kazusa.or.jp/lotus/.
利用表达序列标签(est)、cDNA和基因信息,从日本莲苗mg20基因组文库中筛选出65个具有转化能力的人工染色体(TAC)克隆,并对其核苷酸序列进行了测定。TAC克隆的平均插入长度约为100 kb,本研究测序区域的总长度为6,556,100 bp。加上先前报道的56个TAC克隆的核苷酸序列,目前测序的区域总数为12029295 bp。通过与蛋白质数据库和EST数据库中的序列比对,并通过计算机程序进行基因建模分析,在新测序区域中共鉴定出711个已知或预测功能的潜在蛋白质编码基因,239个基因片段和90个假基因。平均基因密度为1个基因/9140 bp。分配基因的平均长度为2.6 kb,明显大于拟南芥基因组(6451个基因的平均长度为1.9 kb)。在约73%的潜在基因中鉴定出内含子,每个基因平均内含子数和长度分别为3.4 bp和377 bp。根据获得的基因组克隆核苷酸序列,生成简单序列重复长度多态性(SSLP)或衍生的cleaved amplified polymorphic sequence (dCAPS)标记,并利用日本l.l japonicus Gifu B-129和Miyakojima MG-20杂交获得的F2定位群体,将每个克隆定位到连锁图谱上。序列数据、基因信息和图谱信息可通过万维网(http://www.kazusa.or.jp/lotus/)获得。
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引用次数: 37
Biochemical examination of the potential eukaryotic-type protein kinase genes in the complete genome of the unicellular Cyanobacterium synechocystis sp. PCC 6803. 单细胞藻胞杆菌PCC 6803全基因组真核型蛋白激酶基因的生化检测。
A. Kamei, T. Yuasa, Xiaoxing Geng, M. Ikeuchi
The complete genome of the unicellular motile cyanobacterium Synechocystis sp. PCC 6803 harbors seven putative genes for a subfamily Pkn2 of the eukaryotic-type (or "Hanks-type") protein kinase. Previously, SpkA and SpkB were shown to have protein kinase activity and to be required for cell motility. Here, the other five genes were examined. These genes, except for spkG (slr0152), were successfully expressed in Escherichia coli. Eukaryotic-type protein kinase activity of the expressed SpkC (Slr0599), SpkD (S110776) and SpkF (Slr1225) was demonstrated as autophosphorylation and phosphorylation of the general substrate proteins. SpkE (Slr1443) did not show any activity, a finding consistent with its lack of several key amino acid residues in its kinase motif. Gene-disrupted mutants showed no discernible defect in phenotype except that spkD was apparently essential for survival.
单细胞活动蓝藻聚胞杆菌(synnechocystis sp. PCC 6803)的完整基因组包含真核型(或“汉克斯型”)蛋白激酶Pkn2亚家族的七个假定基因。以前,SpkA和SpkB被证明具有蛋白激酶活性,并且是细胞运动所必需的。在这里,其他五个基因被检查。除spkG (slr0152)外,其余基因均在大肠杆菌中成功表达。表达的SpkC (Slr0599), SpkD (S110776)和SpkF (Slr1225)的真核型蛋白激酶活性被证明是自磷酸化和磷酸化一般底物蛋白。SpkE (Slr1443)没有表现出任何活性,这与其激酶基序中缺乏几个关键氨基酸残基一致。基因破坏突变体在表型上没有明显的缺陷,除了spkD显然是生存所必需的。
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引用次数: 42
Conservation of translation initiation sites based on dinucleotide frequency and codon usage in Escherichia coli K-12 (W3110): non-random distribution of A/T-rich sequences immediately upstream of the translation initiation codon. 大肠杆菌K-12 (W3110)中基于二核苷酸频率和密码子使用的翻译起始位点的保护:翻译起始密码子上游的A/ t丰富序列的非随机分布
Kazuo Yamagishi, T. Oshima, Yasushi Masuda, T. Ara, S. Kanaya, H. Mori
Dinucleotide frequencies are useful for characterizing consensus elements as a minimum unit of nucleotide sequence because the neighborhood relations of nucleotide sequences are reflected in dinucleotides. Using a consensus score based on dinucleotide frequencies and intra-species codon usage heterogeneity, denoted by the Z1 parameter, we report the relationship between nucleotide conservation at the translation initiation sites of genes in the Escherichia coli K-12 genome (W3110) and codon usage in its downstream genes. Significant positive correlations were obtained in three regions centered at -13, -4, and +7, which correspond to the Shine-Dalgarno element, the A + T element immediately upstream of the translation initiation site, and the downstream box, respectively.
由于核苷酸序列的邻域关系反映在二核苷酸中,因此二核苷酸频率对于作为核苷酸序列最小单位的一致性元素的特征是有用的。使用基于二核苷酸频率和种内密码子使用异质性(用Z1参数表示)的共识评分,我们报告了大肠杆菌K-12基因组(W3110)中基因翻译起始位点核苷酸保守性与其下游基因密码子使用之间的关系。以-13、-4和+7为中心的3个区域,分别对应翻译起始位点上游的Shine-Dalgarno元件、A + T元件和下游的box,呈显著正相关。
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引用次数: 10
Construction of expression-ready cDNA clones for KIAA genes: manual curation of 330 KIAA cDNA clones (supplement). KIAA基因表达cDNA克隆的构建:330个KIAA cDNA克隆的人工培养(增刊)。
D. Nakajima, N. Okazaki, H. Yamakawa, R. Kikuno, O. Ohara, T. Nagase
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引用次数: 4
Distribution of repetitive sequences on the leading and lagging strands of the Escherichia coli genome: comparative study of Long Direct Repeat (LDR) sequences. 大肠杆菌基因组前后链重复序列的分布:长直接重复序列(LDR)序列的比较研究。
M. Kawano, S. Kanaya, T. Oshima, Yasushi Masuda, T. Ara, H. Mori
In the present study, we developed a method for detecting sequences whose similarity to a target sequence is statistically significant and we examined the distribution of these sequences in the E. coli K-12 genome. Target sequences examined are as follows: (i) short repeat: Crossover hot-spot instigator (Chi) sequence, replication termination (Ter) sequence, and DnaA binding sequence (DnaA box); (ii) potential stem-loop structure repeats: palindromic unit (PU), boxC sequences, and intergenic repeat unit (IRU); (iii) potential RNA coding repeats: rRNAs, PAIR, TRIP, and QUAD; and (iv) potential protein coding repeats: insertion elements (ISs) and Long Direct Repeats (LDRs). We also examined the distribution of these sequences on leading and lagging strands. We obtained another four statistically significant LDR sequences with more than 187 bp matched to LDR-A near the LDR loci, suggesting that these regions might be used as high recombination hot spots for LDR. Adaptation of individual LDRs to E. coli genome is also discussed on the basis of codon usage.
在本研究中,我们开发了一种检测与目标序列相似度具有统计学意义的序列的方法,并检查了这些序列在大肠杆菌K-12基因组中的分布。检测的靶序列如下:(i)短重复序列:交叉热点启动序列(Chi)、复制终止序列(Ter)和dna结合序列(DnaA box);(ii)潜在的茎环结构重复序列:回文单位(PU)、boxC序列和基因间重复单位(IRU);(iii)潜在的RNA编码重复序列:rnas、PAIR、TRIP和QUAD;(iv)潜在的蛋白质编码重复序列:插入元件(ISs)和长直接重复序列(LDRs)。我们还研究了这些序列在前导链和滞后链上的分布。我们在LDR位点附近获得了另外4个具有统计学意义的LDR序列,它们与LDR- a的匹配值超过187bp,这表明这些区域可能是LDR的高重组热点。根据密码子的使用,还讨论了单个ldr对大肠杆菌基因组的适应性。
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引用次数: 11
Characterization of equine microsatellites and microsatellite-linked repetitive elements (eMLREs) by efficient cloning and genotyping methods. 马微卫星和微卫星链接重复元件(eMLREs)的高效克隆和基因分型研究
T. Tozaki, S. Mashima, K. Hirota, N. Miura, N. Choi-Miura, M. Tomita
We performed efficient cloning and genotyping methods for isolation of a large number of polymorphic microsatellites. The methods contain the time-efficient cloning method of constructing microsatellite-enriched libraries and the economic genotyping method of fluorescent labeling of PCR products. Eighty novel equine microsatellites cloned were efficiently isolated from the enrichment library and analyzed for genotype polymorphism. Of these, 72 microsatellites were analyzed with a good resolution. The average heterozygosity of all loci was 0.52, and the number of alleles ranged from one to 9 with an average of 4.5 alleles. The other eight loci showed multiple bands of PCR products, suggesting the occurrence of microsatellites in a repetitive element, in which the number of microsatellite repeats varies among different members of the repetitive element. We found five homologous groups at flanking regions in comparison with the flanking regions of microsatellites from DNA databases. One of them showed homology to equine repetitive element-2. In the other four homologous groups, the two groups were named equine microsatellite-linked repetitive element-1 (eMLRE-1) and equine microsatellite-linked repetitive element-2 (eMLRE-2) as novel equine repetitive elements identified from equine genome. These data should help the analysis of equine DNA sequences and the design of equine genome markers.
我们采用高效的克隆和基因分型方法分离了大量的多态微卫星。这些方法包括构建微卫星富集文库的高效克隆方法和PCR产物荧光标记的经济基因分型方法。从富集文库中高效分离了80个新型马微卫星,并进行了基因型多态性分析。对其中72颗微卫星进行了高分辨率分析。所有位点的平均杂合度为0.52,等位基因数为1 ~ 9个,平均为4.5个。其余8个位点显示多条带PCR产物,提示重复元件中存在微卫星,且微卫星重复数在重复元件的不同成员间存在差异。通过与DNA数据库中微卫星侧翼区域的比较,我们发现了5个同源基团。其中一个与马重复元件-2同源。在其他4个同源类群中,这2个类群分别被命名为马微卫星连锁重复元件-1 (eMLRE-1)和马微卫星连锁重复元件-2 (eMLRE-2),作为从马基因组中鉴定出的新的马重复元件。这些数据将有助于马的DNA序列分析和马基因组标记的设计。
{"title":"Characterization of equine microsatellites and microsatellite-linked repetitive elements (eMLREs) by efficient cloning and genotyping methods.","authors":"T. Tozaki, S. Mashima, K. Hirota, N. Miura, N. Choi-Miura, M. Tomita","doi":"10.1093/DNARES/8.1.33","DOIUrl":"https://doi.org/10.1093/DNARES/8.1.33","url":null,"abstract":"We performed efficient cloning and genotyping methods for isolation of a large number of polymorphic microsatellites. The methods contain the time-efficient cloning method of constructing microsatellite-enriched libraries and the economic genotyping method of fluorescent labeling of PCR products. Eighty novel equine microsatellites cloned were efficiently isolated from the enrichment library and analyzed for genotype polymorphism. Of these, 72 microsatellites were analyzed with a good resolution. The average heterozygosity of all loci was 0.52, and the number of alleles ranged from one to 9 with an average of 4.5 alleles. The other eight loci showed multiple bands of PCR products, suggesting the occurrence of microsatellites in a repetitive element, in which the number of microsatellite repeats varies among different members of the repetitive element. We found five homologous groups at flanking regions in comparison with the flanking regions of microsatellites from DNA databases. One of them showed homology to equine repetitive element-2. In the other four homologous groups, the two groups were named equine microsatellite-linked repetitive element-1 (eMLRE-1) and equine microsatellite-linked repetitive element-2 (eMLRE-2) as novel equine repetitive elements identified from equine genome. These data should help the analysis of equine DNA sequences and the design of equine genome markers.","PeriodicalId":11212,"journal":{"name":"DNA Research: An International Journal for Rapid Publication of Reports on Genes and Genomes","volume":"7 1","pages":"33-45"},"PeriodicalIF":0.0,"publicationDate":"2001-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"85241207","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 40
Structural analysis of a Lotus japonicus genome. I. Sequence features and mapping of fifty-six TAC clones which cover the 5.4 mb regions of the genome. 日本莲基因组的结构分析。1 .覆盖基因组5.4 mb区域的56个TAC克隆的序列特征和图谱。
Shusei Sato, T. Kaneko, Yasukazu Nakamura, E. Asamizu, Tomohiko Kato, S. Tabata
A total of 56 TAC clones with an average insert size of 100 kb were isolated from a TAC library of the Lotus japonicus genome based on the expressed sequences tags (ESTs), cDNA and gene information, and their nucleotide sequences were determined according to the shot-gun based strategy. The total length of the sequenced regions is 5,473,195 bp. By comparison with the sequences in protein and EST databases and analysis with computer programs for gene modeling, a total of 605 potential protein-encoding genes with known or predicted functions, 69 gene segments, and 172 pseudogenes were identified. The average density of the genes assigned so far is 1 gene/8120 bp. Introns were identified in approximately 78% of the potential genes. There was an average of 3.8 introns per gene and the average length of the introns was 375 bp. DNA markers were generated based on the nucleotide sequences obtained, and each clone was mapped onto the linkage map using the F2 mapping population derived from a cross of L. japonicus Gifu B-129 and Miyakojima MG-20. The sequence data, gene information and mapping information are available through the World Wide Web at http://www.kazusa.or.jp/lotus/.
利用表达序列标签(est)、cDNA和基因信息,从日本莲基因组TAC文库中分离出56个平均插入长度为100 kb的TAC克隆,并采用“霰弹枪”策略确定其核苷酸序列。测序区总长度为5,473,195 bp。通过与蛋白质和EST数据库中的序列比对,并结合计算机程序进行基因建模分析,共鉴定出605个已知或预测功能的潜在蛋白质编码基因、69个基因片段和172个假基因。目前分配的基因平均密度为1个基因/8120 bp。在大约78%的潜在基因中鉴定出内含子。每个基因平均含有3.8个内含子,内含子的平均长度为375 bp。利用粳稻Gifu B-129与Miyakojima MG-20杂交获得的F2定位群体,将每个克隆定位到连锁图谱上。序列数据、基因信息和图谱信息可通过万维网(http://www.kazusa.or.jp/lotus/)获得。
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引用次数: 140
Complete genomic sequence of the filamentous nitrogen-fixing cyanobacterium Anabaena sp. strain PCC 7120. 丝状固氮蓝藻Anabaena sp.菌株pcc7120的完整基因组序列。
T. Kaneko, Yasukazu Nakamura, C. Wolk, T. Kuritz, S. Sasamoto, A. Watanabe, Mayumi Iriguchi, Atsuko Ishikawa, K. Kawashima, Takaharu Kimura, Y. Kishida, Mitsuyo Kohara, M. Matsumoto, A. Matsuno, A. Muraki, N. Nakazaki, Sayaka Shimpo, M. Sugimoto, Masaki Takazawa, M. Yamada, Miho Yasuda, S. Tabata
The nucleotide sequence of the entire genome of a filamentous cyanobacterium, Anabaena sp. strain PCC 7120, was determined. The genome of Anabaena consisted of a single chromosome (6,413,771 bp) and six plasmids, designated pCC7120alpha (408,101 bp), pCC7120beta (186,614 bp), pCC7120gamma (101,965 bp), pCC7120delta (55,414 bp), pCC7120epsilon (40,340 bp), and pCC7120zeta (5,584 bp). The chromosome bears 5368 potential protein-encoding genes, four sets of rRNA genes, 48 tRNA genes representing 42 tRNA species, and 4 genes for small structural RNAs. The predicted products of 45% of the potential protein-encoding genes showed sequence similarity to known and predicted proteins of known function, and 27% to translated products of hypothetical genes. The remaining 28% lacked significant similarity to genes for known and predicted proteins in the public DNA databases. More than 60 genes involved in various processes of heterocyst formation and nitrogen fixation were assigned to the chromosome based on their similarity to the reported genes. One hundred and ninety-five genes coding for components of two-component signal transduction systems, nearly 2.5 times as many as those in Synechocystis sp. PCC 6803, were identified on the chromosome. Only 37% of the Anabaena genes showed significant sequence similarity to those of Synechocystis, indicating a high degree of divergence of the gene information between the two cyanobacterial strains.
测定了一株丝状蓝藻Anabaena sp.菌株pcc7120的全基因组核苷酸序列。Anabaena基因组由一条染色体(6413771 bp)和6个质粒组成,分别为pCC7120alpha (408101 bp)、pCC7120beta (186614 bp)、pCC7120gamma (101,965 bp)、pCC7120delta (55,414 bp)、pCC7120epsilon (40,340 bp)和pCC7120zeta (5,584 bp)。染色体包含5368个潜在的蛋白质编码基因,4组rRNA基因,48个tRNA基因代表42个tRNA物种,4个小结构rna基因。45%的潜在蛋白质编码基因的预测产物与已知和预测的已知功能蛋白质的序列相似,27%与假设基因的翻译产物相似。剩下的28%与公共DNA数据库中已知和预测的蛋白质的基因缺乏显著的相似性。根据与报道基因的相似性,60多个参与异囊形成和固氮过程的基因被分配到染色体上。共鉴定出195个编码双组分信号转导系统组分的基因,是Synechocystis sp. PCC 6803的近2.5倍。只有37%的水蓝藻基因序列与聚胞藻基因序列具有显著的相似性,表明两种蓝藻菌株之间的基因信息存在高度差异。
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引用次数: 698
The size and sequence organization of the centromeric region of Arabidopsis thaliana chromosome 4. 拟南芥4号染色体着丝区大小和序列组织。
N. Kumekawa, T. Hosouchi, H. Tsuruoka, Hirokazu Kotani
We have determined the genome structure of the centromeric region of Arabidopsis thaliana chromosome 4 by sequence analysis of BAC clones obtained by genome walking, followed by construction of a physical map using DNA of a hypomethylated strain. The total size of the centromeric region, corresponding to the recombinant inbred (RI) markers between mi87 and mi167, was approximately 5.3 megabases (Mb). This value is over 3 Mb longer than that previously estimated by the Arabidopsis Genome Initiative (Nature, 408, 796-815, 2000). Although we could not cover the entire centromeric region by BAC clones because of the presence of highly repetitive sequences in the middle (2.7 Mb), the cloned regions spanning approximately 1 Mb at both sides of the gap were newly sequenced. These results together with the reported sequences in the adjacent regions suggest that the centromeric region is principally composed of a central domain of 2.7 Mb, consisting of mainly 180-bp repeats and Athila elements, and upper and lower flanking regions of 1.55 Mb and 1 Mb, respectively. The flanking regions were predominantly composed of various types of transposable elements, except for the upper end moiety in which a large 5S rDNA array (0.65 Mb) and central domain-like sequence are present. Such an organization is essentially identical to the centromeric region of chromosome 5 reported previously.
我们通过基因组行走获得的BAC克隆序列分析,确定了拟南芥4号染色体着丝粒区域的基因组结构,然后利用低甲基化菌株的DNA构建了物理图谱。与mi87和mi167之间的重组自交系(RI)标记相对应的着丝粒区总大小约为5.3兆碱基(Mb)。这个值比拟南芥基因组计划(Nature, 408, 796-815, 2000)先前估计的要长3mb以上。虽然我们不能用BAC克隆覆盖整个着丝粒区,因为在中间(2.7 Mb)存在高度重复的序列,但在间隔两侧大约1mb的克隆区域都是新测序的。这些结果和邻近区域的序列表明,着丝粒区主要由2.7 Mb的中心区域和1.55 Mb的上下侧翼区域组成,中心区域主要由180-bp的重复序列和Athila元件组成。侧翼区主要由各种类型的转座元件组成,除了上部部分存在较大的5S rDNA阵列(0.65 Mb)和中心结构域序列。这种组织本质上与先前报道的5号染色体的着丝粒区域相同。
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引用次数: 105
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DNA Research: An International Journal for Rapid Publication of Reports on Genes and Genomes
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