二级出水中抗生素抗性基因存在形态及质粒接合转移影响因素

为解决污水厂二级出水中抗生素抗性基因(antibiotic resistance genes,ARGs)浓度高的问题,深入研究了二级出水中ARGs的相对分子质量分布、主要存在形态,并进一步探究环境因素(时间、温度、pH)对抗性质粒接合转移的影响,为再生水处理工艺研究提供基础.结果 表明:二级出水中的ARGs主要集中在相对分子质量大于100 kDa区间,并且游离态ARGs浓度高于细胞态;抗性质粒pHS-AVC-LW1144可通过在同种属或跨种属细菌间转移扩散来实现接合转移,导致抗性质粒的浓度增加;在0～48 h时间段,抗性质粒浓度呈递增趋势,并在第48小时的浓度最大;低温环境(4℃)和碱性环境(pH=9)更有利于抗性质粒的接合转移.     

细菌遗传转化与水平基因转移

介绍了细菌中水平基因转移、转移途径(转化、接合和转导)以及细菌遗传转化即自然条件中的转化、自然遗传转化及人工转化等研究进展，并且对细菌遗传转化在水平基因转移中的作用进行了探讨．     

蓝藻接合转移系统以及相关因素

蓝藻是一类进行光合放氧的原核生物,蓝藻因其结构的特点,已经成为表达外源基因的理想宿主之一,而接合转移是蓝藻基因转移系统中普遍使用的一种方法。通过接合转移技术可以深入研究光合作用、细胞分化、氮固定和对环境的抵抗。介绍了有关蓝藻接合转移的载体、质粒以及接合转移在蓝藻中的应用,为蓝藻基因工程发展提供信息。     

荚膜红细菌接合转移系统的建立及优化

从培养方式及培养时间对光合细菌Ｒｈｏｄｏｂａｃｔｅｒ ｃａｐｓｕｌａｔｕｓ的ＤＮＡ接合转移系统进行了优化,研究了该接合转移系统的部分特性,发现接合质粒的大小、受体菌细胞膜的特性对接合效率影响较明显,这为光合细菌基因工程菌的构建提供了基础。     

细菌水平基因转移所面临的现实与挑战

水平基因转移对细菌遗传物质进化的影响是存在争议的.本文着重从水平基因转移的进化思想和水平基因转移在进化方面的重要性,特别是在原核细胞进化方面作一综述.首先,阐述水平基因转移在细菌物种进化中的进程;其次,阐述水平基因在不同的进化阶段的范围或广度,此外,讨论重建（或改造）旧的系统发育树关系在面临水平基因转移的可行性;最后,讨论水平基因转移是如何适应新的达尔文进化论学说的,而且需要一种新的进化论去诠释细菌物种的进化机制,水平基因转移就是其中一个很重要的原因.     

规模化鸡场粪样和苍蝇产CMY-2大肠杆菌中耐药基因与毒力基因调查及其共转移研究

为了解全国20家规模化鸡场粪样和苍蝇产CMY-2大肠杆菌中耐药基因与毒力基因的流行现状及其共转移情况,对2013-2015年间分离自粪样和苍蝇中的549株大肠杆菌,采用聚合酶链反应（PCR）方法筛选出blaCMY-2阳性菌株;脉冲场凝胶电泳（PFGE）分析blaCMY-2阳性菌株的遗传进化关系;K-B纸片法检测阳性菌株对13种抗菌药物的耐药性;利用PCR技术检测19种相关耐药基因以及14种毒力基因;接合转移试验和质粒复制子分型研究耐药基因及毒力基因的传播方式。结果,33株大肠杆菌呈CMY-2阳性,阳性率为6.0 %,且均表现为多重耐药;检测到15种耐药基因（blaTEM-1、blaCTX-M-55、blaOXA-1、qnrBDS、aac(6’)-Ib-cr、oqxA、tetA、tetC、sul1、sul2、sul3、rmtB和flor）和4种毒力基因（iutA、traT、fyuA和VagC）;33株携带blaCMY-2基因菌株中有21株接合转移成功,blaCMY-2基因可通过lncA/C或lncI1质粒与耐药基因（blaTEM-1、blaCTX-M-55、tetA或sul2）和（或）毒力基因（traT或VagC）共同转移。本研究阐明了规模化鸡场产CMY-2大肠杆菌携带耐药基因与毒力基因的情况,证实了细菌中质粒介导的耐药与毒力的共转移,为耐药性传播及疾病防控提供了参考。     

转人粒细胞集落刺激因子(hG-CSF)基因的鱼腥藻的构建

本研究目的是构建转G-CSF基因的工程鱼腥藻Anabaena sp..首先将b型G-CSF克隆于中间载体pRL 439和穿梭表达载体pDC-8上.通过三亲接合转移将穿梭表达载体pDCG-CSF转入鱼腥藻内.通过PCR扩增的方法在转基因鱼腥藻中检测到G-CSF基因的存在.     

变构菌素产生菌 S.griseochromogenes 接合转移体系建立与优化

以整合型质粒pSET152为出发质粒,建立并优化了变构菌素产生菌Streptomyces griseochromogenes(S.griseochromogenes)与Escherichia coli(E.coli)的属间接合转移体系,确定了适用于变构菌素产生菌灰产色链霉菌的最佳接合转移条件.研究发现Ca2+和甘氨酸都能够提高接合转移的效率,尤其是甘氨酸的效果非常显著.这一工作对于S.griseochromogenes的基因操作、小分子药物生物合成机制的研究以及具有自主知识产权的药物衍生物的开发具有普遍的应用价值.     

Sanglifehrin A产生菌Streptomyces flaveolus DSM 9954遗传转化系统的建立

Sanglifehrin A的产生菌淡黄色链霉菌Streptomyces flaveolus DSM 9954,通过筛选,得到其菌丝培养基为TSB, 生孢培养基为ISP-4,抗性实验显示淡黄色链霉菌Streptomyces flaveolus DSM 9954对所选抗生素都较敏感. 利用原生质体转化和接合转移都成功地构建了两种遗传转移系统,并且从接合转移成功的链霉菌转化子中回收质粒pKC 1139,经EcoR Ⅰ单酶切电泳,验证了质粒确实通过接合转移从大肠杆菌转移至宿主链霉菌, 这为研究Sangl     

抗砷质粒pSDR941的构建及其在专性自养极端嗜酸性氧化硫硫杆菌中的表达

将具有广泛寄主范围的ＩｎｃＱ质粒ｐＪＲＤ２１５的Ｋｍｒ基因片段删掉，置换进大肠杆菌抗砷质粒ｐＵＭ３上的抗砷基因及Ｐ１启动子片段，构建成新的抗砷质粒ｐＳＤＲ９４１，为１２．４ｋｂ，通过接合的方式将其转移到氧化硫硫杆菌中并获得了表达，与对照菌相比，含质粒ｐＳＤＲ９４１的氧化硫硫杆菌的抗砷水平从１５ｍｍｏｌ／Ｌ提高到３０ｍｍｏｌ／Ｌ．     

Phylogenetic analyses do not support horizontal gene transfers from bacteria to vertebrates.

Horizontal gene transfer (HGT) has long been recognized as a principal force in the evolution of genomes. Genome sequences of Archaea and Bacteria have revealed the existence of genes whose similarity to loci in distantly related organisms is explained most parsimoniously by HGT events. In most multicellular organisms, such genetic fixation can occur only in the germ line. Therefore, it is notable that the publication of the human genome reports 113 incidents of direct HGT between bacteria and vertebrates, without any apparent occurrence in evolutionary intermediates, that is, non-vertebrate eukaryotes. Phylogenetic analysis arguably provides the most objective approach for determining the occurrence and directionality of HGT. Here we report a phylogenetic analysis of 28 proposed HGT genes, whose presence in the human genome had been confirmed by polymerase chain reaction (PCR). The results indicate that most putative HGT genes are present in more anciently derived eukaryotes (many such sequences available in non-vertebrate EST databases) and can be explained in terms of descent through common ancestry. They are, therefore, unlikely to be examples of direct HGT from bacteria to vertebrates.     

Ecology drives a global network of gene exchange connecting the human microbiome

Horizontal gene transfer (HGT), the acquisition of genetic material from non-parental lineages, is known to be important in bacterial evolution. In particular, HGT provides rapid access to genetic innovations, allowing traits such as virulence, antibiotic resistance and xenobiotic metabolism to spread through the human microbiome. Recent anecdotal studies providing snapshots of active gene flow on the human body have highlighted the need to determine the frequency of such recent transfers and the forces that govern these events. Here we report the discovery and characterization of a vast, human-associated network of gene exchange, large enough to directly compare the principal forces shaping HGT. We show that this network of 10,770 unique, recently transferred (more than 99% nucleotide identity) genes found in 2,235 full bacterial genomes, is shaped principally by ecology rather than geography or phylogeny, with most gene exchange occurring between isolates from ecologically similar, but geographically separated, environments. For example, we observe 25-fold more HGT between human-associated bacteria than among ecologically diverse non-human isolates (P = 3.0 × 10(-270)). We show that within the human microbiome this ecological architecture continues across multiple spatial scales, functional classes and ecological niches with transfer further enriched among bacteria that inhabit the same body site, have the same oxygen tolerance or have the same ability to cause disease. This structure offers a window into the molecular traits that define ecological niches, insight that we use to uncover sources of antibiotic resistance and identify genes associated with the pathology of meningitis and other diseases.     

甲基取代基的诱导效应、共轭效应和定位效应

以取代苯苯环碳原子化学位移为依据,用曾经设定并初步证实的甲基诱导效应和共轭效应参数,讨论了甲基取代基的诱导效应、共轭效应及定位效应.化学位移值表示的诱导效应参数表明甲基具有吸电子诱导效应,经校正后的化学位移值表示的共轭效应参数表明甲基具有微弱的给电子共轭效应,甚至其共轭效应有可能正处于给电子和吸电子的临界状态,总之相对于氢原子,甲基具有吸电子效应.甲基有较强的对位定位效应和较弱的邻位定位效应,在适宜条件下间位也有可能发生取代反应.     

Natural conjugative plasmids induce bacterial biofilm development

Horizontal gene transfer is a principal source of evolution leading to change in the ecological character of bacterial species. Bacterial conjugation, which promotes the horizontal transfer of genetic material between donor and recipient cells by physical contact, is a phenomenon of fundamental evolutionary consequence. Although conjugation has been studied primarily in liquid, most natural bacterial populations are found associated with environmental surfaces in complex multispecies communities called biofilms. Biofilms are ideally suited to the exchange of genetic material of various origins, and it has been shown that bacterial conjugation occurs within biofilms. Here I investigate the direct contribution of conjugative plasmids themselves to the capacity of the bacterial host to form a biofilm. Natural conjugative plasmids expressed factors that induced planktonic bacteria to form or enter biofilm communities, which favour the infectious transfer of the plasmid. This general connection between conjugation and biofilms suggests that medically relevant plasmid-bearing strains are more likely to form a biofilm. This may influence both the chances of biofilm-related infection risks and of conjugational spread of virulence factors.     

水平基因转移应用于污染治理的研究进展

随着经济的发展,有毒有害难降解污染物引起的环境问题变得越来越严重。对于这类污染物的处理多采用生化处理方法,其中生物强化技术是目前较好的方法,但是它也存在强化效果差、处理效率低等问题。国外研究人员发现：遗传物质可在不同的生物个体之间或单个细胞的内部细胞器之间自发进行交流,称为水平基因转移。在污染体系中,降解基因的转移能够使土著菌获得降解新功能,从而强化对难降解污染物的处理,这就为现有的生物强化技术提供了一种解决难降解污染问题的新思路。介绍了水平基因转移的概念以及将其应用于污染治理中的研究进展,并分析了技术发展前景。     

引起人非典型性肺炎的冠状病毒基因组与其它冠状病毒基因组的初步比较

一种在世界范围内突然爆发的致命流行病——急性呼吸窘迫综合症(SARS)击倒了数干人，一种全新的冠状病毒被认为是其病原．5个SARS相关冠状病毒的全基因组序列已经完成．我们进行了SARS相关冠状病毒和其它冠状病毒的基因组进化分析和序列比较，结果显示：1．SARS相关冠状病毒不直接来自于任何已知的冠状病毒；2．E蛋白的基因可能是在近期从其它病毒横向转移到SARS病毒中来的；3．Sl和S2基因发生了较大范围的缺失或插入突变．这些基因横向转移和突变改变了SARS相关病毒的表面结构和抗原性，极有可能是导致其获得侵染人类细胞的主要原因．     

The bacterial conjugation protein TrwB resembles ring helicases and F1-ATPase

The transfer of DNA across membranes and between cells is a central biological process; however, its molecular mechanism remains unknown. In prokaryotes, trans-membrane passage by bacterial conjugation, is the main route for horizontal gene transfer. It is the means for rapid acquisition of new genetic information, including antibiotic resistance by pathogens. Trans-kingdom gene transfer from bacteria to plants or fungi and even bacterial sporulation are special cases of conjugation. An integral membrane DNA-binding protein, called TrwB in the Escherichia coli R388 conjugative system, is essential for the conjugation process. This large multimeric protein is responsible for recruiting the relaxosome DNA-protein complex, and participates in the transfer of a single DNA strand during cell mating. Here we report the three-dimensional structure of a soluble variant of TrwB. The molecule consists of two domains: a nucleotide-binding domain of alpha/beta topology, reminiscent of RecA and DNA ring helicases, and an all-alpha domain. Six equivalent protein monomers associate to form an almost spherical quaternary structure that is strikingly similar to F1-ATPase. A central channel, 20 A in width, traverses the hexamer.     

基于增强步态能量图和Gabor特征的辨别共同向量身份识别

提出了一种通过统计行走模式的动态信息进行步态识别方法.对代表每一类的步态能量图像(GEI)进行方差分析,以求得动态权值掩模(DWM).通过DWM对原始GEI进行动态和形状信息的增强,以获得新的步态表征EGEI.为增加可辨识信息,使用一组Gabor小波对EGEI进行卷积,然后采用辨别共同向量分析(DCV)将高维卷积结果在低维空间表示.通过使用简单的分类策略在USF步态数据库上的对比实验,证明了本方法对识别性能提高的有效性.     

基于分子轨道隐式溶剂H2O下苏氨酸的电荷转移

基于分子轨道（MO）和自然跃迁轨道（NTO）成分计算分子片段间的电荷转移. 先用密度泛函理论(DFT)中的CAM-B3LYP方法, 在6-31G(d)基组水平上优化隐式溶剂H₂O下苏氨酸（Thr）分子的几何构型,  再在相同理论方法下进行含时密度泛函理论(TDDFT)的电子激发计算, 给出隐式溶剂H₂O下Thr分子体系电子激发过程中片段间电荷转移特征的对比结果. 结果表明： 在S1~S5激发态中, 仅S2中有一对MO32→MO33跃迁轨道占绝对优势, 可通过分析该轨道的成分讨论电荷转移; 其他激发态可通过NTO分析方法讨论电荷转移; S0向激发态S1,S3和S4电荷转移的主要贡献为NTO32→NTO33轨道, 与Hirshfeld和Becke方法的定性结果一致, 定量结果略有差别.