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1.
Some have hypothesized that ancestral proteins were, on average, less specific than their descendants. If true, this would provide a universal axis along which to organize protein evolution and suggests that reconstructed ancestral proteins may be uniquely powerful tools for protein engineering. Ancestral sequence reconstruction studies are one line of evidence used to support this hypothesis. Previously, we performed such a study, investigating the evolution of peptide-binding specificity for the paralogs S100A5 and S100A6. The modern proteins appeared more specific than their last common ancestor (ancA5/A6), as each paralog bound a subset of the peptides bound by ancA5/A6. In this study, we revisit this transition, using quantitative phage display to measure the interactions of 30,533 random peptides with human S100A5, S100A6, and ancA5/A6. This unbiased screen reveals a different picture. While S100A5 and S100A6 do indeed bind to a subset of the peptides recognized by ancA5/A6, they also acquired new peptide partners outside of the set recognized by ancA5/A6. Our previous work showed that ancA5/A6 had lower specificity than its descendants when measured against biological targets; our new work shows that ancA5/A6 has similar specificity to the modern proteins when measured against a random set of peptide targets. This demonstrates that altered biological specificity does not necessarily indicate altered intrinsic specificity, and sounds a cautionary note for using ancestral reconstruction studies with biological targets as a means to infer global evolutionary trends in specificity.  相似文献   
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Bio3D is a family of R packages for the analysis of biomolecular sequence, structure, and dynamics. Major functionality includes biomolecular database searching and retrieval, sequence and structure conservation analysis, ensemble normal mode analysis, protein structure and correlation network analysis, principal component, and related multivariate analysis methods. Here, we review recent package developments, including a new underlying segregation into separate packages for distinct analysis, and introduce a new method for structure analysis named ensemble difference distance matrix analysis (eDDM). The eDDM approach calculates and compares atomic distance matrices across large sets of homologous atomic structures to help identify the residue wise determinants underlying specific functional processes. An eDDM workflow is detailed along with an example application to a large protein family. As a new member of the Bio3D family, the Bio3D‐eddm package supports both experimental and theoretical simulation‐generated structures, is integrated with other methods for dissecting sequence‐structure–function relationships, and can be used in a highly automated and reproducible manner. Bio3D is distributed as an integrated set of platform independent open source R packages available from: http://thegrantlab.org/bio3d/ .  相似文献   
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Ants (Hymenoptera: Formicidae) are dominant social insects that play important ecological roles in terrestrial ecosystems. Camponotus leydigi (Forel) is widely distributed in the Neotropical region and is frequently found in the Brazilian cerrado savanna interacting with plants and other insects. Field observations indicate that C. leydigi has a polydomous nesting habit, but little is known about the genetic relationship among workers. In this study, we identify the first nine microsatellite loci for C. leydigi that will allow further investigation on its genetic diversity. We used a microsatellite-enriched library method. According to this method, repetitive sequences are captured with (CT)8 and (GT)8 biotin-linked probes, with subsequent recovery by streptavidin magnetic-coated beads. We observed that eight loci were polymorphic. The mean (± standard error) observed and expected heterozygosities were 0.55 ± 0.23 and 0.73 ± 0.28, respectively. The rarified allelic richness ranged from 1 to 5.32. The polymorphism contents were similar to diversity estimates found in markers previously developed for other Camponotus ants. These markers will be useful for future studies on population genetics and ecology of Camponotus ants in cerrado, including nesting ecology, colony structure, dispersal and conservation.  相似文献   
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随着流感病毒基因组测序数据的急剧增加,深入挖掘流感病毒基因组大数据蕴含的生物学信息成为研究热点。基于中国流感病毒流行特征数据,建设一个集自动化、一体化和信息化的序列库系统,对于实现流感病毒基因组批量快速翻译、注释、存储、查询、分析具有重要的应用价值。本课题组通过集成一系列软件和工具包,并结合自主研发的其他功能,在底层维护的2个关键的参考数据集基础上另外追加了翻译注释信息最佳匹配的精细化筛选规则,构建具有流感病毒基因组信息存储、自动化翻译、蛋白序列精准注释、同源序列比对和进化树分析等功能的自动化系统。结果显示,通过Web端输入fasta格式的流感病毒基因序列,本系统可针对参考序列片段数据集(blastdb.fasta)进行Blast同源性检索,可以鉴定流感病毒的型别(A、B或C)、亚型和基因片段(1~8片段);在此基础上,通过查询数据库底层用于翻译、注释的基因片段参考数据集,可以获得一组肽段数据集,然后通过循环调用ProSplign软件对其进行预测。结合精细化的筛选准入规则,选出与输入序列匹配最好的翻译后产物,作为该输入序列的预测蛋白,输出为gbk,asn和fasta等通用格式的文件,给出序列长度、是否全长、病毒型别、亚型、片段等信息。基于以上工作,另外自主研发了系统其他的附加功能如进化树分析展示、基因组数据存储等功能,构建成基于Web服务的流感病毒基因组自动化翻译注释系统。本研究提示,系统高度集成系列软件以及自有的注释翻译数据库文件,实现从序列存储、翻译、注释到序列分析和展示的功能,可全面满足我国高通量基因检测数据共享化、本土化、一体化、自动化的需求。  相似文献   
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急性出血性结膜炎(Acute hemorrhagic conjunctivitis,AHC)是目前人类最常见的眼病之一,柯萨奇病毒A组24型变异株(Coxsackievirus A24 variant,CV-A24v)是近年来报道引起该病的主要病原体。本研究选取10株来自江西省2010年AHC暴发疫情的CV-A24v,采用特异性引物扩增并测定其全基因组序列。对该10条CV-A24v的全基因组序列进行系统发育分析以及重组分析,计算本研究测定的江西10条以及GenBank中所有22条CV-A24v的全基因组序列的氨基酸置换熵值,并预测其正向选择位点。结果表明,在江西10条CV-A24v基因组序列中未检测到重组。基于全基因组序列构建的最大似然树表明江西10株CV-A24v属于GIV基因型,且分处于两条传播链。对上述32条CV-A24v序列的氨基酸置换熵值计算,共得到25个易突变位点(熵值>0.6),易突变概率最高的区段为2A区。基于Datamonkey中FUBAR和FEL模型分析,发现位于结构蛋白VP2区的234位氨基酸为两种模型共同获得的CV-A24v的正向选择位点。本研究分析了江西10株CV-A24v的全基因组序列特征,为CV-A24v引起的AHC防控工作提供了基础资料。  相似文献   
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Since the first revelation of proteins functioning as macromolecular machines through their three dimensional structures, researchers have been intrigued by the marvelous ways the biochemical processes are carried out by proteins. The aspiration to understand protein structures has fueled extensive efforts across different scientific disciplines. In recent years, it has been demonstrated that proteins with new functionality or shapes can be designed via structure-based modeling methods, and the design strategies have combined all available information — but largely piece-by-piece — from sequence derived statistics to the detailed atomic-level modeling of chemical interactions. Despite the significant progress, incorporating data-derived approaches through the use of deep learning methods can be a game changer. In this review, we summarize current progress, compare the arc of developing the deep learning approaches with the conventional methods, and describe the motivation and concepts behind current strategies that may lead to potential future opportunities.  相似文献   
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为构建灰盖拟鬼伞Coprinopsis cinerea的核定位蛋白重组表达系统,本研究通过蛋白序列比对和信息学分析,预测了灰盖拟鬼伞组蛋白H2B的核定位序列,构建了融合组蛋白H2B核定位序列的绿色荧光蛋白(green fluorescent protein,GFP)重组表达载体,将该载体转入灰盖拟鬼伞AmutBmut菌株,检测组蛋白H2B核定位序列是否可以引导GFP定位在细胞核区域。研究结果显示,在灰盖拟鬼伞β-微管蛋白启动子的驱动下,经过密码子优化的GFP基因可以在鬼伞体内表达、产生绿色荧光。同时来自灰盖拟鬼伞组蛋白H2Bb (CC1G_07639)的核定位序列,可以成功地引导GFP入核。本研究成功地构建了灰盖拟鬼伞核定位蛋白重组表达系统,为后续优化灰盖拟鬼伞的CRISPR/Cas9基因编辑系统、深入研究灰盖拟鬼伞的子实体发育机制奠定了基础。  相似文献   
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BackgroundsDiverse marine habitats along Jeddah's Red Sea coast support rich biodiversity. Few studies have been done on its diverse communities, especially its microbial counterparts. Metagenomic analysis of marine benthic micro-eukaryotic communities was performed for the first time on the Red Sea coast of Jeddah. This research looks into their community structure and metabolic potential.MethodsNext-generation sequencing was used to examine the micro-eukaryotic communities of seven sedimentary soil samples from four Jeddah coast locations. After isolating DNA from seven benthic sedimentary soil samples, the 18S rDNA V4 regions were amplified and sequenced on the Illumina MiSeq. It was also verified using an Agilent Technologies 2100 Bioanalyzer with a DNA 1000 chip (Agilent Technologies, Fisher Scientific). A standard curve of fluorescence readings generated by qPCR quantification using the Illumina library was achieved using the GS FLX library. Metagenomic data analysis was used to evaluate the microbial communities' biochemical and enzymatic allocations in studied samples.ResultsBlast analysis showed that the top ten phyla were Annelida, Eukaryota, Diatomea, Porifera, Phragmoplastophyta, Arthropoda, Dinoflagellata, Xenacoelomorpha Nematoda, and uncultured. Annelida was also found in the highest percentage (93%), in the sample M followed by Porifera (64%), the most abundant in the control sample then Eukaryotes (61%), Phragmatoplastophyta (55%), Arthropoda, and Diatomea (the least common) (32%). community diversity analysis: using Shannon and inverse Simpson indices showed sediment composition to be effective. Also, PICRUST2 indicated that the most abundant pathways were pyruvate fermentation to isobutanol, pyrimidine deoxyribonucleotide phosphorylation, adenosine ribonucleotide de novo biosynthesis, guanosine ribonucleotide de novo biosynthesis, NAD salvage pathway I, the super pathway of glyoxylate bypass and aerobic respiration I (cytochrome c).ConclusionResults showed that high throughput metagenomics could reveal species diversity and estimate gene profiles. Environmental factors appear to be more important than geographic variation in determining the structure of these microbial communities. This study provides the first report of marine benthic micro-eukaryotic communities found on the Red Sea coast of Jeddah and will serve as a good platform for future research.  相似文献   
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