recombination

重组
  • 文章类型: Journal Article
    第一代多倍体通常比已建立的野生多倍体种群遭受更多的减数分裂错误和更低的生育力。一个这样的例子是起源于c的拟南芥的异源多倍体模型物种。16000代以前。我们在这里使用细胞学和基因组方法的组合来比较自然进化和第一代“合成”的减数分裂及其结果。我们表明,虽然自然系中的减数分裂很大程度上是二倍体样的,合成品系具有高水平的减数分裂错误,包括不完全突触和非同源交叉形成。后代的全基因组重新测序显示,源自合成亲本的同源交换水平高20倍,非整倍性高8倍。同源交换表现出强烈的远端偏向,主要发生在基因中,定期产生新的蛋白质变体。我们还观察到,当发生在反向同态区域时,同源交换可以产生大碱基级别的INDEL。最后,我们观察到的证据表明,在多倍体适应性方面存在性别差异,当使用合成植物作为母本时,与天然品系的相互杂交成功率更高.我们的结果直接联系了A的细胞学表型和它们的基因组结果,证明同源杂交是新异源多倍体基因组不稳定的基础,并且比同源杂交更偏向远端。
    First-generation polyploids often suffer from more meiotic errors and lower fertility than established wild polyploid populations. One such example is the allopolyploid model species Arabidopsis suecica which originated c. 16 000 generations ago. We present here a comparison of meiosis and its outcomes in naturally evolved and first-generation \'synthetic\' A. suecica using a combination of cytological and genomic approaches. We show that while meiosis in natural lines is largely diploid-like, synthetic lines have high levels of meiotic errors including incomplete synapsis and nonhomologous crossover formation. Whole-genome re-sequencing of progeny revealed 20-fold higher levels of homoeologous exchange and eightfold higher aneuploidy originating from synthetic parents. Homoeologous exchanges showed a strong distal bias and occurred predominantly in genes, regularly generating novel protein variants. We also observed that homoeologous exchanges can generate megabase scale INDELs when occurring in regions of inverted synteny. Finally, we observed evidence of sex-specific differences in adaptation to polyploidy with higher success in reciprocal crosses to natural lines when synthetic plants were used as the female parent. Our results directly link cytological phenotypes in A. suecica with their genomic outcomes, demonstrating that homoeologous crossovers underlie genomic instability in neo-allopolyploids and are more distally biased than homologous crossovers.
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  • 文章类型: Journal Article
    经济地理学家如何确定从哪里开始他们的研究项目,在哪里找到和界定他们的案例研究,在哪里以及如何“切入”问题?在缺乏对这些问题的不言而喻或预先给出的答案的情况下,从哪里开始以及如何开始的问题和选择不可避免地与初步概念化甚至理论化的问题纠缠在一起,因为案件与其说是发现的,不如说是发现的,以各种方式与不同的“理论-方法包”共同生产。“对于这些问题,没有(也可以)单一或普遍的答案。相反,这个简短的干预概述了“开始”的一个理由,“这样的理论基础应该参考一种特定的理论方法或模式。这里的方法集中在重组开发的问题上,关于扩展案例研究设计的作用,以及仍然稀疏地意识到的结合分析模式的潜力。
    How do economic geographers determine where to begin their research projects, where to locate and delimit their case studies, where and how to \"cut in\" to problems? In the absence of self-evident or pregiven answers to these questions, the problem-cum-choice of where and how to start is inescapably tangled up with issues of preliminary conceptualization and indeed theorization, since cases are not so much found as made, being in various ways coproduced with different \"theory-method packages.\" There is (and can be) no singular or universal answer to these questions. Instead, this brief intervention outlines one rationale for getting \"started,\" founded as such rationales should be with reference a particular approach or mode of theorization. The approach here centers on the problematic of recombinant development, on the role of extended case-study designs, and on the still sparsely realized potential of conjunctural modes of analysis.
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  • 文章类型: Journal Article
    减数分裂重组由Po11产生的DNA双链断裂(DSB)引发,Po11是一种II型拓扑异构酶样蛋白,与DSB末端共价连接。虽然已经在几种生物中检测到了Spo11寡核苷酸-Mre11的核解去除产物,共价Spo11-DSB前体的寿命尚未确定,并且可以进行替代处理。这里,我们探索了人酪氨酸DNA磷酸二酯酶的活性,TDP2-一种已知修复DNA末端的蛋白质,其来自分离自酿酒酵母细胞的Po11DSB上的失效拓扑异构酶活性。我们证明,即使在存在竞争基因组DNA的情况下,TDP2也可以从ssDNA寡核苷酸和dsDNA末端去除Sp11肽。有趣的是,TDP2处理的DSB末端难以被Exo1切除,这表明Mre11产生的ssDNA在体内可能是必需的,以促进Spo11DSB处的HR,即使TDP2具有活性。此外,虽然TDP2可以在体外去除Po11肽,减数分裂细胞中的TDP2表达无法在体内去除Spo11,这与它帮助修复拓扑异构酶诱导的DNA损伤的能力相反。这些结果表明,spo11-DNA,但不是拓扑异构酶-DNA切割复合物,TDP2酶无法进入,可能是由于减数分裂重组位点的高阶蛋白质复合物闭塞所致。
    Meiotic recombination is initiated by DNA double-strand breaks (DSBs) created by Spo11, a type-II topoisomerase-like protein that becomes covalently linked to DSB ends. Whilst Spo11 oligos-the products of nucleolytic removal by Mre11-have been detected in several organisms, the lifetime of the covalent Spo11-DSB precursor has not been determined and may be subject to alternative processing. Here, we explore the activity of human Tyrosyl DNA Phosphodiesterase, TDP2-a protein known to repair DNA ends arising from abortive topoisomerase activity-on Spo11 DSBs isolated from S. cerevisiae cells. We demonstrate that TDP2 can remove Spo11 peptides from ssDNA oligos and dsDNA ends even in the presence of competitor genomic DNA. Interestingly, TDP2-processed DSB ends are refractory to resection by Exo1, suggesting that ssDNA generated by Mre11 may be essential in vivo to facilitate HR at Spo11 DSBs even if TDP2 were active. Moreover, although TDP2 can remove Spo11 peptides in vitro, TDP2 expression in meiotic cells was unable to remove Spo11 in vivo-contrasting its ability to aid repair of topoisomerase-induced DNA lesions. These results suggest that Spo11-DNA, but not topoisomerase-DNA cleavage complexes, are inaccessible to the TDP2 enzyme, perhaps due to occlusion by higher-order protein complexes at sites of meiotic recombination.
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  • 文章类型: Journal Article
    生殖系细胞产生配子,是有性生殖所必需的特化细胞.生殖系细胞首先通过几轮有丝分裂扩增,然后切换到减数分裂程序,这需要特定的蛋白质组进行DNA重组,染色体配对,和隔离。令人惊讶的是,我们以前发现突触复合体的一些蛋白质,前期I减数分裂结构,在果蝇雌性的有丝分裂区域中已经表达和需要。这里,为了评估其他减数分裂基因是否比预期更早表达,我们分离了有丝分裂和减数分裂细胞群,以比较它们的RNA含量。我们的转录组学分析表明,所有已知的减数分裂I基因已经在有丝分裂区表达;然而,只有一部分是翻译的。作为一个案例研究,我们专注于Po11的果蝇同源物mei-W68,以评估其在mRNA和蛋白质水平上的表达,并使用不同的突变等位基因来测定减数分裂前的功能。在分裂的生殖细胞中,我们无法检测到Mei-W68在同源染色体配对中的任何功能作用。我们的研究为在有丝分裂区域表达的减数分裂基因的进一步功能分析铺平了道路。
    Germline cells produce gametes, which are specialized cells essential for sexual reproduction. Germline cells first amplify through several rounds of mitosis before switching to the meiotic program, which requires specific sets of proteins for DNA recombination, chromosome pairing, and segregation. Surprisingly, we previously found that some proteins of the synaptonemal complex, a prophase I meiotic structure, are already expressed and required in the mitotic region of Drosophila females. Here, to assess if additional meiotic genes were expressed earlier than expected, we isolated mitotic and meiotic cell populations to compare their RNA content. Our transcriptomic analysis reveals that all known meiosis I genes are already expressed in the mitotic region; however, only some of them are translated. As a case study, we focused on mei-W68, the Drosophila homolog of Spo11, to assess its expression at both the mRNA and protein levels and used different mutant alleles to assay for a premeiotic function. We could not detect any functional role for Mei-W68 during homologous chromosome pairing in dividing germ cells. Our study paves the way for further functional analysis of meiotic genes expressed in the mitotic region.
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  • 文章类型: Journal Article
    重组通过将现有变体重新改组为新的遗传背景来破坏遗传连锁。传统上,这些动态是通过检查等位基因之间的相关性来量化的,以及它们如何作为重组率的函数衰减。然而,这些相关性的大小受到自然选择和遗传漂移等其他进化力量的强烈影响,这使得很难梳理出重组的影响。在这里,我们介绍了一个理论框架,用于分析另一个统计族,该统计族测量了重组产生的同质。我们得出了分析表达式,可以预测这些统计数据如何取决于重组和复发突变率,负选择和遗传漂移的强度,和突变等位基因的当前频率。我们发现同质的程度可以很大程度上取决于这个频率范围,这反映了这些突变发生的潜在时间尺度。我们展示了如何使用这些缩放特性来隔离重组的影响,并讨论它们对细菌水平基因转移速率的影响。
    Recombination breaks down genetic linkage by reshuffling existing variants onto new genetic backgrounds. These dynamics are traditionally quantified by examining the correlations between alleles, and how they decay as a function of the recombination rate. However, the magnitudes of these correlations are strongly influenced by other evolutionary forces like natural selection and genetic drift, making it difficult to tease out the effects of recombination. Here we introduce a theoretical framework for analyzing an alternative family of statistics that measure the homoplasy produced by recombination. We derive analytical expressions that predict how these statistics depend on the rates of recombination and recurrent mutation, the strength of negative selection and genetic drift, and the present-day frequencies of the mutant alleles. We find that the degree of homoplasy can strongly depend on this frequency scale, which reflects the underlying timescales over which these mutations occurred. We show how these scaling properties can be used to isolate the effects of recombination, and discuss their implications for the rates of horizontal gene transfer in bacteria.
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  • 文章类型: Journal Article
    星状病毒是单链的,能够感染人类以及广泛的哺乳动物和鸟类物种的正义RNA病毒,长度约为6.6-7.7kb。在这项研究中,从广西采集的139份山羊粪便样本进行RT-PCR检测,其中两个是山羊星状病毒阳性,阳性率为1.44%(2/139)。星状病毒株的全基因组序列和星状病毒株的部分基因组序列,命名为GXWZ2023和GXHC2023,进行了扩增和测序,它们的序列长度分别为6284nt和6213nt,分别。其中,山羊星状病毒GXHC2023的衣壳蛋白与绵羊星状病毒GX的氨基酸同一性最高,为95.9%,属于MAstV-2基因型。然而,发现GXWZ2023株的最近亲是山羊星状病毒四川,核苷酸序列同一性为76.8%。该菌株的ORF1ab非结构蛋白与绵羊星状病毒S5.1和山羊星状病毒G5.1菌株的氨基酸同一性最高,分别为89.2和95.8%,分别。然而,其ORF2衣壳蛋白与牛星状病毒(BAstV)162021CHN菌株具有68.4%的氨基酸同一性,与所有可用的山羊星状病毒菌株仅具有21.9-64%的氨基酸同一性。GXWZ2023菌株在ORF2地区与中国(BAstV162021CHN)和日本牛菌株(BAstVJPN2015)重组。因此,根据国际病毒分类学委员会的物种分类标准,山羊星状病毒GXWZ2023被提议为山羊星状病毒科的新成员。这些发现增强了我们对山羊星状病毒的流行和遗传进化的理解,并为将来在其他动物中研究这些病毒提供了科学依据。
    Astroviruses are single-stranded, positive-sense RNA viruses capable of infecting humans as well as a wide range of mammalian and avian species, with a length of approximately 6.6-7.7 kb. In this study, 139 goat fecal samples collected from the Guangxi province were used for the RT-PCR detection, and two of these were positive for goat astrovirus, with a positivity rate of 1.44% (2/139). The complete genome sequence of an astrovirus strain and the partial genome sequence of a strain astrovirus, named GX WZ 2023 and GX HC 2023, were amplified and sequenced, and their sequence lengths were 6284 nt and 6213 nt, respectively. Among them, the capsid protein of goat astrovirus GX HC 2023 showed the highest amino acid identity of 95.9% with ovine astrovirus GX, which belonged to the MAstV-2 genotype. However, the closest relative of the GX WZ 2023 strain was found to be the caprine astrovirus Sichuan, with a nucleotide sequence identity of 76.8%. The ORF1ab nonstructural protein of this strain showed the highest amino acid identities of 89.2 and 95.8% with the ovine astrovirus S5.1 and caprine astrovirus G5.1 strains, respectively. However, its ORF2 capsid protein has 68.4% amino acid identity with the bovine astrovirus (BAstV) 16 2021 CHN strain and only 21.9-64% amino acid identity with all available strains of goat astrovirus. The GX WZ 2023 strain was recombined with the Chinese (BAstV 16 2021 CHN) and Japanese bovine strains (BAstV JPN 2015) in the ORF2 region. Therefore, the goat astrovirus GX WZ 2023 is proposed as a new member of the family goat astroviridae based on the species classification criteria of the International Committee on Taxonomy of Viruses. These findings enhance our understanding of the prevalence and genetic evolution of goat astrovirus and provide a scientific basis for future studies of these viruses in other animals.
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  • 文章类型: Journal Article
    检测,表征,SARS-CoV-2重组变体的监测对全球公共卫生当局构成了挑战。重组变体,由两个或多个SARS-CoV-2谱系组成,通常对传播有未知的影响,免疫逃逸,和出现早期的毒力。我们检查了2020年至2022年在加利福尼亚州收集的4213个SARS-CoV-2重组SARS-CoV-2基因组,以描述区域和全州的流行趋势。许多这些重组基因组,例如属于XZ谱系或新的重组谱系的那些,可能起源于加利福尼亚州。我们讨论了围绕潘戈血统分配的挑战和局限性,使用公开的序列数据,和足够的样本大小进行流行病学分析。尽管随着全球SARS-CoV-2测序量的减少,这些挑战仍将继续,这项研究增强了我们迄今为止对SARS-CoV-2重组基因组的理解,同时为未来对新兴重组谱系的认识奠定了基础.
    The detection, characterization, and monitoring of SARS-CoV-2 recombinant variants constitute a challenge for public health authorities worldwide. Recombinant variants, composed of two or more SARS-CoV-2 lineages, often have unknown impacts on transmission, immune escape, and virulence in the early stages of emergence. We examined 4213 SARS-CoV-2 recombinant SARS-CoV-2 genomes collected between 2020 and 2022 in California to describe regional and statewide trends in prevalence. Many of these recombinant genomes, such as those belonging to the XZ lineage or novel recombinant lineages, likely originated within the state of California. We discuss the challenges and limitations surrounding Pango lineage assignments, the use of publicly available sequence data, and adequate sample sizes for epidemiologic analyses. Although these challenges will continue as SARS-CoV-2 sequencing volumes decrease globally, this study enhances our understanding of SARS-CoV-2 recombinant genomes to date while providing a foundation for future insights into emerging recombinant lineages.
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  • 文章类型: Journal Article
    真菌遗传系统理想地结合了用于基因组操作的分子工具和有性生殖系统,以创建基因组修饰组合的信息分类。当利用性周期来产生多突变体时,亲本的背景基因型变异可能导致后代表型变异,掩盖了联合突变的影响。这里,为了减轻轮枝镰刀菌的这种变异,我们产生的MAT1-2菌株与测序的野生型MAT1-1菌株接近同基因,FGSC7600。这是通过将FGSC7600与发散的野生型MAT1-2菌株FGSC7603杂交,然后进行六次连续回交来实现的(例如,六代)MAT1-2后代为FGSC7600。我们对每一代进行测序并绘制重组事件图。亲本杂交涉及11条染色体中的9条染色体上的26条杂交。在FGSC7603中发现但在FGSC7600中缺乏的可有可有可无的12号染色体在第5代后代中不存在。经常观察到没有交叉的完整染色体的遗传。大约140千碱基的缺失,在4号染色体上包含54个预测基因,发生在第4代,并保留在第5代,这表明这些基因对于生长以及无性和有性生殖都是可有可无的。最终的MAT1-2菌株TMRU10/35与FGSC7600约93%相同。TMRU10/35可从真菌遗传学储备中心获得FGSC27326,从ARS培养物收集中心获得NRRL64809。
    Fungal genetic systems ideally combine molecular tools for genome manipulation and a sexual reproduction system to create an informative assortment of combinations of genomic modifications. When employing the sexual cycle to generate multi-mutants, the background genotype variations in the parents may result in progeny phenotypic variation obscuring the effects of combined mutations. Here, to mitigate this variation in Fusarium verticillioides, we generated a MAT1-2 strain that was near isogenic to the sequenced wild-type MAT1-1 strain, FGSC7600. This was accomplished by crossing FGSC7600 with the divergent wild-type MAT1-2 strain FGSC7603 followed by six sequential backcrosses (e.g., six generations) of MAT1-2 progeny to FGSC7600. We sequenced each generation and mapped recombination events. The parental cross involved twenty-six crossovers on nine of the eleven chromosomes. The dispensable chromosome 12, found in FGSC7603 but lacking in FGSC7600, was not present in the progeny post generation five. Inheritance of complete chromosomes without crossover was frequently observed. A deletion of approximately 140 kilobases, containing 54 predicted genes on chromosome 4, occurred in generation 4 and was retained in generation 5 indicating that these genes are dispensable for growth and both asexual and sexual reproduction. The final MAT1-2 strain TMRU10/35 is about 93% identical to FGSC7600. TMRU10/35 is available from the Fungal Genetics Stock Center as FGSC27326 and from the ARS Culture Collection as NRRL64809.
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  • 文章类型: Journal Article
    猪繁殖与呼吸综合征(PRRS)是全球养猪业的重大威胁,它在泰国的流行超过二十年。
    为了解泰国PRRS病毒(PRRSV)GP5基因的遗传变异和重组,我们从NCBI数据库中检索到726个GP5基因序列。系统发育树使用邻居连接(NJ)和最大似然(ML)方法构建,并进行重组分析。
    对83株PRRSV-1和83株PRRSV-2进行同源性分析。系统发育分析显示PRRSV-1和PRRSV-2株在泰国流行,后者表现出更广泛的分布。PRRSV-1菌株聚集在进化枝A中,D,H,而PRRSV-2菌株分为谱系1、5和亚谱系8.7,进一步分为8.7/HP和8.7/NA亚谱系。子系8.7/NA菌株占循环PRRSV-2菌株的很大比例。同源性分析显示,PRRSV-1的核苷酸和氨基酸相似性范围为75.4至100.0%和41.3至100.0%,PRRSV-2的78.6至100.0%和70.8至100.0%。氨基酸序列比对显示突变,插入,和PRRSV-1GP5中的缺失,以及与生物学功能相关的PRRSV-2GP5中的关键残基突变。重组分析鉴定了PRRSV-2亚谱系8.7菌株内的两个重组事件。
    这些发现证实了GP5蛋白的变异性。这项研究增强了我们对泰国PRRSV流行和遗传变异的理解,为PRRS预防和控制提供有价值的见解。
    UNASSIGNED: Porcine reproductive and respiratory syndrome (PRRS) is a significant threat to the global swine industry, and its prevalence in Thailand spans over two decades.
    UNASSIGNED: To understand the genetic variation and recombination of the PRRS virus (PRRSV) GP5 gene in Thailand, we retrieved 726 GP5 gene sequences from the NCBI database. Phylogenetic trees were constructed using the neighbor-joining (NJ) and maximum likelihood (ML) methods, and recombination analysis was performed.
    UNASSIGNED: Homology analysis was conducted on 83 PRRSV-1 and 83 PRRSV-2 strains. Phylogenetic analysis revealed the prevalence of both PRRSV-1 and PRRSV-2 strains in Thailand, with the latter exhibiting wider distribution. PRRSV-1 strains clustered into clades A, D, and H, while PRRSV-2 strains grouped into lineages 1, 5, and sublineage 8.7, further divided into 8.7/HP and 8.7/NA sublineages. Sublineage 8.7/NA strains accounted for a significant proportion of circulating PRRSV-2 strains. Homology analysis showed nucleotide and amino acid similarities ranging from 75.4 to 100.0% and 41.3 to 100.0% for PRRSV-1, and 78.6 to 100.0% and 70.8 to 100.0% for PRRSV-2 strains. Amino acid sequence alignments revealed mutations, insertions, and deletions in PRRSV-1 GP5, and key residue mutations in PRRSV-2 GP5 associated with biological functions. Recombination analysis identified two recombination events within PRRSV-2 sublineage 8.7 strains.
    UNASSIGNED: These findings confirm the variability of the GP5 protein. This study enhances our understanding of PRRSV prevalence and genetic variation in Thailand, contributing valuable insights for PRRS prevention and control.
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  • 文章类型: Journal Article
    减数分裂是产生配子的发育程序。为了产生健康的配子,减数分裂重组会在每对同源染色体之间产生相互交换,从而促进忠实的染色体分离。利用裂变酵母和生化,遗传,和细胞学方法,我们研究了CDK(细胞周期蛋白依赖性激酶)在Swi5-Sfr1控制中的作用,Swi5-Sfr1是一种Rad51重组酶辅助因子,参与重组过程中同源物的侵袭。我们证明Sfr1是CDK目标,其磷酸化在减数分裂前期下调Swi5-Sfr1功能。磷酸化模拟sfr1-7D突变体的表达抑制Rad51结合,它强大的染色体负载,并随后减少同源物之间的重组。另一方面,非磷酸化sfr1-7A突变体在后期改变Rad51动力学,并加剧了在dbl2缺失突变体中观察到的染色质分离缺陷和Rad51保留。我们提出Sfr1磷酸抑制作为一种新的细胞周期依赖性机制,这确保了重组中间体的及时解决和成功的染色体分配到配子中。此外,Sfr1的N端无序部分,一个进化保守的特征,作为协调这种磷酸调节的监管平台,蛋白质定位和稳定性,几个CDK位点和调控序列是保守的。
    Meiosis is the developmental program that generates gametes. To produce healthy gametes, meiotic recombination creates reciprocal exchanges between each pair of homologous chromosomes that facilitate faithful chromosome segregation. Using fission yeast and biochemical, genetic, and cytological approaches, we have studied the role of CDK (cyclin-dependent kinase) in the control of Swi5-Sfr1, a Rad51-recombinase auxiliary factor involved in homolog invasion during recombination. We show that Sfr1 is a CDK target, and its phosphorylation downregulates Swi5-Sfr1 function in the meiotic prophase. Expression of a phospho-mimetic sfr1-7D mutant inhibits Rad51 binding, its robust chromosome loading, and subsequently decreases interhomolog recombination. On the other hand, the non-phosphorylatable sfr1-7A mutant alters Rad51 dynamics at late prophase, and exacerbates chromatin segregation defects and Rad51 retention observed in dbl2 deletion mutants when combined with them. We propose Sfr1 phospho-inhibition as a novel cell-cycle-dependent mechanism, which ensures timely resolution of recombination intermediates and successful chromosome distribution into the gametes. Furthermore, the N-terminal disordered part of Sfr1, an evolutionarily conserved feature, serves as a regulatory platform coordinating this phospho-regulation, protein localization and stability, with several CDK sites and regulatory sequences being conserved.
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