中华鳖4个Sox基因保守区的序列分析

采用PCR技术,扩增和克隆了中华鳖Sox基因（TSSox）。经DNA序列分析显示,Sox基因在系统进化上十分保守,其中TSSox4与鸟类LF4基因编码的氨基酸序列完全相同、与人类SOX4和Sox4编码的序列仅一个氨基酸的差异;TSSox5与鸟类的LF5基因的编码也仅一个氨基酸发生了改变;TSSox2与海龟的TSox2相似性最高。4条TSSox序列中,TSSox与人SRY基因序列相似性最高,达75％;序列上的相似性可能暗示了它们在功能上的保守性。     

互补克隆法:利用酵母鉴定油菜基因

克隆植物基因的一个重要方法是利用另一种生物的等效基因。不同真核生物间的基因杂交仅限于亲缘关系很近的品种。动物探针与植物DNA的杂交也仅限于在进化过程中高度保守的基因。另一种可能就是用异源抗体筛选表位的表达文库。该技术的问题在于保守性表位因其在被免疫动物中的保守性而常常是非致免疫的。基因互补是依据两种生物基因间的功能相似性而非序列相似性发展起来的第三种技术。其基本思想是制备一种生物的基因序列文库并将之插入缺乏某种基因功能的另一种生物中。在第二种生物中由第一种生     

斑马鱼Gfi-1 基因结构和功能分析

目的:研究斑马鱼Gfi-1基因在物种间进化的保守性和功能分析。方法:运用生物信息学方法分析斑马鱼Gfi-1基因结构特征和保守性等。结果:斑马鱼Gfi-1基因在蛋白水平与小鼠、人高度保守;分析斑马鱼和人的Gfi-1基因外显子、内含子和ATG起始、终止密码子也具有高度相似性;从进化树分析斑马鱼Gfi-1基因与人、小鼠、犬、猴等在进化上高度保守;分析斑马鱼、人、小鼠Gfi-1基因在染色体上的位置和相邻基因,显示出惊人的相似性。结论:斑马鱼Gfi-1基因在进化上高度保守,为脊椎动物保守基因,为其后续在造血系统和造血微环境方面的研究提供了理论支持和铺垫。     

hsa-miR-192-3p的靶基因预测及功能分析

通过生物信息学方法预测hsa-miR-192-3p的靶基因及其靶基因的可能功能。首先通过miRbase在线数据库对hsamiR-192-3p的碱基序列及序列在各物种间的保守性进行分析,再通过miRGator v3. 0在线数据库查看hsa-miR-192-3p在各个组织器官中的表达丰富度情况;其次,应用Target Scan和miRanda在线数据库预测hsa-miR-192-3p的靶基因;最后,将预测得到的两个数据库的靶基因交集用DAVID在线数据库进行功能富集分析和信号转导通路富集分析。结果表明:hsa-miR-192-3p在人、家鼠、猕猴等生物中存在高度保守性; hsa-miR-192-3p在胃肠道、肾脏、肝胆系统、干细胞、鼻、脾、胸腺中表达丰富度较高;通过两个靶基因预测软件预测的靶基因取交集后共有190个;功能富集分析发现hsa-miR-192-3p靶基因富集在细胞质、细胞核、质膜、高尔基体等15个细胞组件(p0. 05),参与蛋白结合、GTP酶活性、锌离子跨膜转运蛋白活性等7个分子功能(p0. 05),涉及金属离子运输、RNA聚合酶II启动子的转录阳性调控、基因表达调节、钙离子跨膜运输、胚胎发育等18个生物过程(p0. 05);预测靶基因集合显著富集于癌症通路与催乳素信号通路中(p0. 05)。得出结论:hsa-miR-192-3p预测的靶基因集合富集于多个生物过程,与肿瘤密切相关,生物信息预测为今后的研究奠定了一定的理论基础,为后续实验验证提供了研究方向。     

肝癌细胞HepG2中增强子的识别及生物信息学分析

目的:整合增强子特征识别肝癌细胞HepG2增强子,并对其保守性、GC含量、转录因子调控、靶基因功能等进行分析,以期解析肝癌细胞增强子参与的调控网络。方法:通过整合H3K27ac、H3K4me1和H3K4me3组蛋白修饰及DNaseⅠ高敏位点的Chip-seq数据预测HepG2中的增强子,计算每个增强子的平均Phast Cons分数和GC含量,评估整体增强子的保守性与GC含量,整合ENCODE转录因子结合位点数据寻找转录因子-增强子调控,使用GREAT和DAVID分别对增强子和增强子的靶基因进行GO与KEGG通路功能富集分析。结果:共识别2254个肝细胞癌增强子,1432个增强子靶基因,135个转录因子的9983个增强子结合位点;比较随机位点靶基因,发现增强子显著正调控靶基因的表达;保守性与GC含量分析表明增强子具有显著高的保守性与GC含量,并存在C-T/C-T/C-T-G模式的motif;增强子功能分析显示增强子显著富集于蛋白结合、酶结合、转录因子结合、RNA聚合酶Ⅱ结合等已知增强子功能,增强子GO与KEGG通路功能富集分析表明增强子靶基因显著参与细胞增殖、细胞凋亡、细胞周期调控和细胞迁移等肿瘤相关的生物进程与信号通路。结论:识别的肝细胞癌增强子具有显著高的保守性与GC含量,受多种转录因子调控,对其靶基因起正调控作用并且显著富集于肿瘤相关生物学进程与信号通路中。     

hsa-miR-381-3p靶基因预测及其相关信号通路的生物信息学分析

应用生物信息学方法分析miR-381-3p序列,预测其靶基因,并对靶基因进行蛋白质互作分析、功能富集分析及信号转导通路富集分析。结果发现,已知的成熟miR-381-3p序列在不同物种间高度保守。蛋白质互作分析显示,mi R-381-3p预测靶基因所编码蛋白质间存在复杂的相互作用,尤其是靶基因BTBD1、NUP160、STX16等编码的蛋白质,在互作中起关键作用。GO分析发现其靶基因集合可能参与细胞过程、生物调节、细胞大分子代谢等生物学过程;KEGG通路分析发现其靶基因集合显著富集于mTOR、Wnt、p53、MAPK等信号通路中。分析结果初步提示miR-381-3p通过调控靶基因广泛参与多种重要的生物学过程,为后续的实验性研究提供了线索。     

miR-15a靶基因的预测及生物信息学分析

目的:对目前研究较为广泛的miR-15a的靶基因进行预测及相关生物信息学分析,以期为miR-15a靶基因的实验验证提供数据支持,并为深入研究miR-15a的调控机制及生物学功能奠定基础和提供理论指导。方法:选择TargetScan5.1与PicTar两种计算方法预测miR-15a的靶基因的交集作为分析的基因集合,分别进行GO注释描述、GO富集分析和生物通路富集分析。结果与结论:预测靶基因集合分别富集在转录调控、蛋白质修饰、细胞周期等生物学过程和蛋白激酶活性等分子功能上(P0.01);经典miR-15a预测靶基因集合显著富集于KEGG通路数据库中的Wnt信号通路、细胞周期和p53信号通路等5个信号转导通路及前列腺癌、慢性髓细胞性白血病、黑素瘤等7个疾病通路中(P0.05)。     

中华绒螯蟹Dmrt基因保守区段的克隆和序列分析

Dmrt基因家族是一个与性别决定和发育相关的基因家族,在不同进化类型的生物中具有相当的保守性。采用PCR技术,扩增了中华绒螯蟹Dmrt基因的DM结构域。经序列分析,获得了Dmrt基因家族的4个成员基因EsDmrt2a、EsDmrt2b、EsDmrt2c和EsDmrt5。结果表明,不同进化地位动物的Dmrt2和Dmrt2基因DM域编码序列存在高度的同源性,显示Dmrt基因在系统进化上高度保守,序列上的相似性可能暗示着它们在功能上的保守性。     

大绿蛙Sox基因和Dmrt基因的序列分析

Sox基因家族与Dmrt基因家族在胚胎发育及性别分化中起着重要作用。采用PCR方法,扩增了大绿蛙Sox基因和Dmrt基因的保守区,分别获得长约220 bp和140 bp的片段。序列分析表明,大绿蛙雌雄个体之间Sox基因、Dmrt基因序列没有差异,与人和其它动物的Sox基因、Dmrt基因有非常高的相似性,显示了Sox基因及Dmrt基因在系统进化上的高度保守性。本研究为探讨大绿蛙的性别决定机制及Sox基因与Dmrt基因的进化提供了分子资料。     

miR-335在肿瘤组织中的表达及其预测靶基因的生物信息学分析

目的:分析miR-335在多种肿瘤组织与癌旁组织中的表达,预测其靶基因并进行相关生物信息学分析,为进一步研究miR-335在肿瘤中的调控机制提供理论基础。方法:分析miR-335的保守性及在多个肿瘤组织中的表达;预测miR-335靶基因,并使用DAIVID对miR-335靶基因进行生物信息学分析。结果:miR-335序列高度保守,在肝癌、肺癌、乳腺癌、肝内胆管癌、脂肪肉瘤中表达下调(P<0.05)。预测miR-335靶基因共34个,靶基因集合功能富集于细胞迁移、凋亡、转录调控,以及蛋白质分子连接、细胞骨架组成等生物学过程和分子功能(P<0.05);主要参与了轴突向导和黏着斑信号通路、黑素瘤疾病信号通路及TGF-β信号通路(P<0.05)。结论:miR-335在多种肿瘤中表达异常,且涉及多个生物学过程和信号转导通路,与肿瘤的发生发展密切相关。     

Conservation unit allows assessing vulnerability and setting conservation priorities for a Mediterranean endemic plant within the context of extreme urbanization

Current biodiversity patterns are a temporary state in a continuum of ecological and evolutionary changes. Conservation policies must incorporate this dynamic to ensure the long-term conservation of biodiversity which is particularly challenging in a context of extreme urbanization. An original approach, rarely used for plant conservation, is to define conservation units to set conservation priorities within species by combining ecological and evolutionary divergences. In the Maritime Alps (southern France), the ecological and evolutionary divergences between the populations of Acis nicaeensis has allowed us the determination of conservation units of this endemic plant threatened by severe urbanization and land-use changes. Phylogeographical data (cpDNA haplotype) were considered as a proxy for evolutionary legacy, and ecological data (multivariate analysis of habitat) were used as a proxy for ecological distinctiveness. Our goal was to explore the potential of this approach to assess vulnerability and set conservation priorities for narrow endemic species in the context of conflict between biodiversity and human activities. The results highlight five different conservation units within A. nicaeensis distribution. Genetic and ecological divergences are present at fine-scale. This pattern is highly endangered by urbanisation. This study highlights the vulnerability of conservation units near the coast whose originality and restricted distribution call for rapid conservation management to avoid the loss of species evolutionary and ecological distinctiveness.     

Fine-scale spatial genetic structure and gene flow in a small, fragmented population of Sinojackia rehderiana (Styracaceae), an endangered tree species endemic to China

Populations of Sinojackia rehderiana are highly threatened and have small and scattered distribution due to habitat fragmentation and human activities. Understanding changes in genetic diversity, the fine-scale spatial genetic structure (SGS) at different life stages and gene flow of S. rehderiana is critical for developing successful conservation strategies for fragmented populations of this endangered species. In this study, 208 adults, 114 juveniles and 136 seedlings in a 50 × 100-m transect within an old-growth forest were mapped and genotyped using eight microsatellite makers to investigate the genetic diversity and SGS of this species. No significant differences in genetic diversity among different life-history stages were found. However, a significant heterozygote deficiency in adults and seedlings may result from substantial biparental inbreeding. Significant fine-scale spatial structure was found in different life-history stages within 19 m, suggesting that seed dispersal mainly occurred near a mother tree. Both historical and contemporary estimates of gene flow (13.06 and 16.77 m) indicated short-distance gene dispersal in isolated populations of S. rehderiana. The consistent spatial structure revealed in different life stages is most likely the result of limited gene flow. Our results have important implications for conservation of extant populations of S. rehderiana. Measures for promoting pollen flow should be taken for in situ conservation. The presence of a SGS in fragmented populations implies that seeds for ex situ conservation should be collected from trees at least 19-m apart to reduce genetic similarity between neighbouring individuals.     

Determination and validation of principal gene products

MOTIVATION: Alternative splicing has the potential to generate a wide range of protein isoforms. For many computational applications and for experimental research, it is important to be able to concentrate on the isoform that retains the core biological function. For many genes this is far from clear. RESULTS: We have combined five methods into a pipeline that allows us to detect the principal variant for a gene. Most of the methods were based on conservation between species, at the level of both gene and protein. The five methods used were the conservation of exonic structure, the detection of non-neutral evolution, the conservation of functional residues, the existence of a known protein structure and the abundance of vertebrate orthologues. The pipeline was able to determine a principal isoform for 83% of a set of well-annotated genes with multiple variants.     

Consequences of near and far between-population crosses for offspring fitness in a rare herb

Crosses between plants from different populations may result in heterosis or outbreeding depression. However, despite its importance for conservation, little is known about the spatial scale over which these effects may arise. To investigate the consequences of between-population crosses at two distinct spatial scales, we conducted reciprocal crosses between four populations from two regions in the rare perennial herb Aster amellus . We assessed seed set and offspring fitness in a common garden experiment. Overall, between-population crosses within regions (10 km) resulted in 8% lower seed set than within-population crosses, while between-region crosses (70 km) resulted in 17% higher seed set than within-population crosses. Moreover, offspring from between-population crosses produced 18% more flower heads than offspring from within-population crosses. We conclude that hybridisation between A. amellus plants from different populations did not lead to immediate outbreeding depression and, thus, could represent a valid conservation option to increase genetic diversity. Moreover, our results suggest that the distance between populations affects the outputs of between-population crosses and therefore needs to be taken into account when promoting gene flow between populations.     

Exploring the effectiveness of integrated conservation and development interventions in a Central African forest landscape

Integrated conservation and development projects (ICDPs) have had limited success in addressing the often conflicting objectives of conservation and development. We developed a model with local participants to explore the trade-offs between conservation and development in southeastern Cameroon, where illegal hunting is regarded as the greatest challenge to conservation. We simulated the effects of different ICDP strategies by varying the degree of focus on antipoaching activities, anticorruption measures and direct development investments, and by varying the overall budget for such activities. Our outcome variables were numbers of selected wildlife species and household incomes. The model outcomes from the different scenarios were used to stimulate debate among stakeholders. Contributing to poverty alleviation while maintaining current animal population sizes will be extremely difficult and will require long-term external financial support. Devoting greater attention to improving local environmental governance emerged as the highest priority for this investment. We used the model outputs to inform some of the major policy makers in the region. Participatory modeling is a valuable means of capturing the complexities of achieving conservation at landscape scales and of stimulating innovative solutions to entrenched problems.     

The role of taxonomy in species conservation

Taxonomy and species conservation are often assumed to be completely interdependent activities. However, a shortage of taxonomic information and skills, and confusion over where the limits to 'species' should be set, both cause problems for conservationists. There is no simple solution because species lists used for conservation planning (e.g. threatened species, species richness estimates, species covered by legislation) are often also used to determine which units should be the focus of conservation actions; this despite the fact that the two processes have such different goals and information needs. Species conservation needs two kinds of taxonomic solution: (i) a set of practical rules to standardize the species units included on lists; and (ii) an approach to the units chosen for conservation recovery planning which recognizes the dynamic nature of natural systems and the differences from the units in listing processes that result. These solutions are well within our grasp but require a new kind of collaboration among conservation biologists, taxonomists and legislators, as well as an increased resource of taxonomists with relevant and high-quality skills.     

What works for threatened species recovery? An empirical evaluation for Australia

Despite the growing numbers of threatened species and high levels of spending on their recovery worldwide, there is surprisingly little evidence about which conservation approaches are effective in arresting or reversing threatened species declines. Using two government data sets, we examined associations between population trends for 841 nationally-threatened terrestrial species in Australia, and four measures of conservation effort: (a) how much their distribution overlaps with strictly protected areas (IUCN I–IV), (b) and other protected areas (IUCN V–VI), (c) the number of recovery activities directed at the species, and (d) numbers of natural resource conservation activities applied in areas where populations of the threatened species occur. We found that all populations of 606 (72%) species were in decline. Species with greater distributional overlap with strictly protected areas had proportionately more populations that were increasing or stable. This effect was robust to geographic range size, data quality differences and extent of protection. Measures other than strictly protected areas showed no positive associations with stable or increasing trends. Indeed, species from regions with more natural resource conservation activities were found to be more likely to be declining, consistent with differential targeting of such generalised conservation activities to highly disturbed landscapes. Major differences in trends were also found among the different jurisdictions in which species predominantly occurred, which may be related to different legislative protections against habitat destruction. Although we were not able to test causation, this research corroborates other evidence that protected areas contribute to the stabilization or recovery of threatened species, and provides little empirical support for other conservation approaches.     

Ant community structure during forest succession in a subtropical forest in South-East China

Understanding how communities respond to environmental gradients is critical to predict responses of species to changing habitat conditions such as in regenerating secondary habitats after human land use. In this study, ground-living ants were sampled with pitfall traps in 27 plots in a heterogeneous and diverse subtropical forest to test if and how a broad set of environmental variables including elevation, successional age, and tree species richness influence ant diversity and community composition. In total, 13,441 ant individuals belonging to 71 species were found. Ant abundance was unrelated to all environmental variables. Rarefied ant species richness was negatively related to elevation, and Shannon diversity decreased with shrub cover. There was considerable variation in ant species amongst plots, associated with elevation, successional age, and variables related to succession such as shrub cover. It is shown that younger secondary forests may support a species-rich and diverse community of ants in subtropical forests even though the species composition between younger and older forests is markedly different. These findings confirm the conservation value of secondary subtropical forests, which is critical because subtropical forests have been heavily exploited by human activities globally. However, the findings also confirm that old-growth forest should have priority in conservation as it supports a distinct ant community. Our study identifies a set of ant species which are associated with successional age and may thus potentially assist local conservation planning.     

Policy indicators for use in impact evaluations of protected area networks

The number of protected areas (PAs) has steadily increased in the past 20 years, but their effectiveness to meet conservation targets is consistently questioned. Most conservation impact evaluations of protected areas assume that formal designations, like that of IUCN categories, reflect site-specific conservation rules, but this is not always true. In this paper we illustrate how conservation rules could be empirically assessed by use of content analysis combined with optimal scaling. This flexible methodology allows us to quantitatively assess strictness levels for use in conservation impact evaluations. The strictness measures could also indicate whether conservation rules are consistently applied in the different IUCN categories thereby providing guidance for future assignment of PAs to the IUCN protected area management categories. We illustrate how policy indicators based on conservation rules could be developed in two contrasting mountain protected area networks in Norway and in British Columbia (BC), including a total of 48 PAs in Norway and 51 in BC. Conservation rules for recreational use, motorized access and resource use were quantitatively assessed, thus providing a measure of how strictly PAs regulate the different human activities. Our results show that the main differences in strictness are between the two countries, followed by the contrast between national parks and provincial parks in BC. Overall, Norway has a more liberal conservation policy than BC and older national parks in BC have a much stricter conservation policy than most of the other PAs in this study. Overarching conservation objectives did not reflect the level of strictness (the conservation rules) that guide the daily management of individual PAs. This applies to both countries. We recommend to empirically investigate site-specific conservation rules to include de facto management of human activities in conservation policy impact evaluations. The methodology is also useful for monitoring downgrading of the protected area status, which is a result of authorizing human activities that are not consistent with conservation objectives.