环境微生物宏基因组学研究中的生物信息学方法

高通量测序技术的发展促进了组学技术在环境微生物研究中的广泛应用,而宏基因组学是目前最为关键和成熟的组学方法。生物信息学在微生物宏基因组学研究中具有至关重要的作用。它贯穿于宏基因组学的数据收集和存储、数据处理和分析等各个阶段,既是宏基因组学推广的最大瓶颈,也是目前宏基因组学研究发展的关键所在。本文主要介绍和归纳了目前在高通量宏基因组测序中常用的生物信息学分析平台及其重要的信息分析工具。未来几年之内,测序成本的下降和测序深度的增加将进一步增大宏基因组学研究在数据存储、数据处理和数据挖掘层面的难度,因此相应生物信息学技术与方法的研究和发展也势在必行。近期内我们应该首先加强基础性分析和存储平台的建设以方便普通环境微生物研究者使用,同时针对目前生物信息分析的瓶颈步骤和关键任务重点突破,逐步发展。     

Genome-wide identification of antimicrobial peptides in the liver fluke,Clonorchis sinensis

The increase in prevalence of antimicrobial resistance makes the search for new antibiotic agents imperative. Antimicrobial peptides (AMPs) from natural resources have been recognized as suitable tools to combat antibiotic-resistant bacteria. The liver fluke Clonorchis sinensis living in germ-filled environments could be a good source of antimicrobials. Here, we report the use of a rational protocol that combines AMP predictions based on their physicochemical properties and their in vivo stability to discover AMP candidates from the entire genome of C. sinensis. To screen AMP candidates, in silico analyses based on the physicochemical properties of known AMPs, such as length, charge, isoelectric point, and in vitro and in vivo aggregation values were performed. To enhance their in vivo stability, proteins having proteolytic cleavage sites were excluded. As a consequence, four high-activity, highstability peptides were identified. These peptides could be potential starting materials for the development of new AMPs via structural modification and optimization. Thus, this study proposes a refined computational method to develop new AMPs and identifies four AMP candidates, which could serve as templates for further development of peptide antibiotics.     

棉花抗逆相关基因GhDr1的克隆及生物信息学分析

通过TAIL-PCR的方法从棉花中克隆到未知功能的新基因GhDr1,生物信息学分析结果表明,GhDr1的预测蛋白含有酪氨酸激酶和蛋白酶C磷酸化位点、潜在的锌指类功能模块、MAPK磷酸化位点及MAPK相互作用识别位点;与GhDr1同源基因位于同一基因簇的基因多为参与逆境信号转导的蛋白。推测GhDr1基因可能在逆境信号转导途径的磷酸化级联反应中被调节,参与棉花逆境应答的生理过程。     

麻疯树鲨烯合酶基因SQS的克隆及生物信息学分析

以麻疯树(Jatropha curcas L.)总RNA为模板,根据已报道的鲨烯合酶基因序列设计简并引物,用RACE方法克隆得到麻疯树鲨烯合酶基因全长cDNA,命名为JcSQS。JcSQS全长1609 bp,包含1个1242 bp的开放阅读框,预测麻疯树鲨烯合酶基因编码的蛋白含有413个氨基酸。JcSQS具有鲨烯合酶类的保守结构域,JcSQS 蛋白与蓖麻、柿、木榄等植物中SQS基因编码的氨基酸序列具有高度同源性。这为研究麻疯树萜烯类物质的生物合成和调控机制奠定了基础。     

基于EST数据库进行SNP分子标记开发的研究进展及在猕猴桃属植物中的应用研究

对基于EST数据库开发SNP标记的特点、开发策略等进行了综述,并介绍了在中华猕猴桃复合体(Actinidia chinesis Planch.)中开发EST-SNP的基本思路和初步结果,为后续分子实验验证及其在自然居群中的应用奠定基础,并为其它相关物种的EST-SNP分子标记开发提供借鉴.     

牡丹PsCS基因的克隆及序列分析

以牡丹品种‘赵粉’(Paeonia suffruticosa L.cv.‘Zhao Fen’)为试材,采用RT-PCR和RACE方法从雄蕊中获得了一个牡丹柠檬酸合成醇(citrate synthase,CS)基因cDNA全长,命名为PsCS,GenBank登录号为HQ449568.其cDNA全长1 564 bp,包含75 bp的5’非编码区、73 bp的3 '非编码区和一个长度为1 416 bp编码471个氨基酸的开放阅读框.序列比对和系统进化分析表明,PsCS与葡萄的亲缘关系最近,相似性达89.4％以上.     

蜜蜂TPI基因克隆与生物信息学预测

利用电子克隆方法获得蜜蜂(Apis mellifera)磷酸甘油醛异构酶(triosephosphate isomerase,TPI)基因,并采用生物信息学方法对该基因编码蛋白从等电点、疏水性/亲水性、二级结构等进行了预测,以及试验验证,结果表明蜜蜂TPI基因全长为1 768 bp,具有完整的开放阅读框架(ORF),并得到了试验证实.     

The real cost of sequencing: higher than you think!

Advances in sequencing technology have led to a sharp decrease in the cost of ''data generation''. But is this sufficient to ensure cost-effective and efficient ''knowledge generation''?     

The role of statistical power analysis in quantitative proteomics

Designing an experiment for quantitative proteomic analysis is not a trivial task. One of the key factors influencing the success of such studies is the number of biological replicates included in the analysis. This, along with the measured variation will determine the statistical power of the analysis. Presented is a simple yet powerful analysis to determine the appropriate sample size required for reliable and reproducible results, based on the total variation (technical and biological). This approach can also be applied retrospectively for the interpretation of results as it takes into account both significance (p value) and quantitative difference (fold change) of the results.