甲基营养型酵母表达载体系统研究进展

酵母作为生产真核异源蛋白的宿主菌 ,具备了易于分子遗传操作 ,原核微生物的生长性质及真核蛋白的翻译后加工修饰等特点 .其中Ｓａｃｃｈａｒｏｍｙｃｅｓｃｅｒｅｖｉｓｉａｅ (酿酒酵母 )是第一个作为异源蛋白基因表达系统并加以应用的酵母 .人们认为Ｓ .ｃｅｒｅｖｉｓｉａｅ并非表达外源基因的最理想的酵母宿主菌 .在一群优于Ｓ .ｃｅｒｅｖｉｓｉａｅ的表达菌中 ,甲基营养型酵母Ｐｉｃｈｉａｐａｓｔｏｒｉｓ研究得最多 ,最具应用潜力 .在 2 0世纪 70年代期间 ,Ｐ .ｐａｓｔｏｒｉｓ是单细胞蛋白 (Ｓｉｎｇｌｅ ｃｅｌｌｐｒｏｔｅｉｎ…     

鸡贫血病毒的分子生物学研究进展

鸡贫血病毒 (ｃｈｉｃｋｅｎａｎｅｍｉａｖｉｒｕｓ,ＣＡＶ)是鸡传染性贫血病 (ｃｈｉｃｋｅｎｉｎｆｅｃｔｉｏｕｓａｎｅｍｉａ,ＣＩＡ)的主要病原 .ＣＡＶ感染可引起雏鸡骨髓造血组织和胸腺等淋巴组织萎缩 ,从而导致严重贫血 ,甚至造成死亡[1] ;同时ＣＡＶ感染还能导致免疫抑制[2～ 7] .自 1979年日本学者Ｙｕａｓａ等[8] 首次报道该病毒以来 ,西德、瑞典、英国、丹麦、波兰、美国、澳大利亚、荷兰、巴西和阿根廷等国先后发现该病毒的存在[9～ 11] .我国于 1992年首次在黑龙江省分离到ＣＡＶ[12 ] ,随后在河南、山东、江苏、辽宁、吉林…     

群感效应与信号分子在污泥颗粒化过程中的作用研究进展

好氧颗粒污泥具有良好的沉降性能和较好的污水处理效果等优点.本文综述好氧颗粒污泥的形成过程,重点分析在污泥颗粒化过程中群感效应和细胞信号分子的作用.研究发现,群感效应和细胞信号分子在污泥颗粒化过程中发挥着重要作用,群感效应可以加快污泥胞外聚合物合成和提高微生物表面附着能力,信号分子可以促进微生物分泌胞外聚合物并影响生物膜的形成.举例说明信号分子中的c-di-GMP作为第二信使在污泥颗粒化过程中的重要作用,c-di-GMP可以将细菌从单细胞游走型转变为多细胞黏附型——生物膜状态.投加钙离子会促进能够分泌c-diGMP的菌群生长从而加快好氧颗粒污泥的形成,投加锰离子会减少好氧颗粒污泥中c-di-GMP的含量导致好氧颗粒污泥解体.最后进行前景展望,认为从群感效应和信号分子的角度去解析好氧颗粒污泥形成机理,可为确立好氧颗粒污泥形成机制奠定理论基础,对其进一步的工程化应用具有现实意义.     

藻际环境铁载体细菌的筛选及群体感应信号对其合成的调节作用

铁载体是海洋微生物获取铁元素的重要代谢因子，现有的一些研究推测海洋微生物铁代谢功能的发挥依赖群体效应，但海洋微生物群体行为的介导方式及其对铁载体功能的调节还未见揭示.为深入了解微生物的铁代谢特征，以藻际微生物为例，从中分离筛选产铁载体的细菌，并探讨群体感应信号（QS）对其合成的调控作用.利用稀释涂布法从甲藻赤潮样品中分离得到5株产铁载体的细菌，16Sr RNA基因序列鉴定为新鞘氨醇杆菌（Novosphingobium sp.）、鞘氨醇单胞菌（Sphingomonas sp.）、寡养单胞菌（Stenotrophomonas sp.）、嗜水气单胞菌（Aeromonas hydr ophi la）和根瘤菌（R hizobiumar enae）.铬天青S(CAS)检测法结合化学显色反应分析显示，细菌分泌的铁载体主要包括羧酸盐、异羟肟酸、儿茶酚和肠杆菌素.薄层色谱法（TLC）检测到4株细菌具有明显产群体感应信号的能力，信号分子主要为高丝氨酸内酯（AHLs）AHL-C8、AHL-C8-O以及AHL-10.为验证群体感应信号对铁载体的调节能力，以新鞘氨醇杆菌为代表构建AHL突变株，通过突变株与野生株...     

光合细菌光合产氢机理研究进展[综述]

７０年代以来，随着全球性的能源危机和温室效应的加剧，无污染、可再生的氢能的研究开发日受重视．物制氢技术是氢能开发研究的一项重要内容．至今已知的具产氢活性的微生物有“光合细菌”（ｐｈｏｔｏｓｙｎｔｈｅｔｉｃｂａｃｔｅｒｉａ，ＰＳＢ）、藻类（ａｌｇａｅ）和非光合细菌（ｎｏｎｐｈｏｔｏｓｙｎｔｈｅｔｉｃｂａｃｔｅｒｉａ）［１］．由于ＰＳＢ光合产氢的速度要比藻类快，能量利用率比非光合细菌高，且能将产氢与光能利用、有机物的去除有机地耦合在一起，因而得到了众多研究者的关注．本文就ＰＳＢ的产氢的机理及影响产…     

动胶菌属系统分类、生理特征及其在活性污泥中的作用

微生物菌群一直是活性污泥研究过程中的关键问题,近年来通过分子生物学等研究手段已经发现动胶菌是活性污泥微生物菌群中的优势细菌.本文以活性污泥微生物菌群为出发点,重点梳理了动胶菌属分类学研究历史和现状,并建议在前期命名过程中出现的垂(悬)丝状动胶菌和生枝动胶菌ATCC 19623株及ATCC 25935株不应再作为动胶菌属细菌对待.同时介绍了动胶菌属细菌的基本特性及其在活性污泥法处理废水中降解有机物、除磷脱氮和吸附重金属离子等方面的应用,以及动胶菌胞外多聚物的组成和动胶菌优良菌株的选育.我们及合作者最近分析了两个喜树脂动胶菌的基因组并鉴定出参与菌胶团形成的一个大型基因簇,也实现了基因定点敲除等遗传操作.今后研究应重点关注动胶菌分子遗传学、功能基因组学及胞外多聚物生物合成和菌胶团形成机理、化学结构和性质.     

信号分子对类球红细菌(Rhodobacter sphaeroides)碳源利用的影响

类球红细菌拥有cer群体感应系统.它既有细胞间信息传递的功能,也对碳源利用进行调节.为调查自诱导物对碳源利用的影响,在仅少量EDTA作唯一能源物的培养基中加入N-(tetradecanoyl)-DL-homoserine lactone,通过测定活菌数变化观察这种类球红细菌自诱导物的结构类似物对碳源利用的影响.然后利用Biolog-GN鉴定板,在加入外源信号分子和对照的条件下,研究了类球红细菌正常培养与饥饿培养下对95种单一碳源的不同利用情况.结果表明,在存在信号分子的饥饿情况下,细菌能更好的利用EDTA维持活菌数量.当培养到6h的时候,加入信号分子培养的鉴定板中,正常培养情况下细菌对碳源的利用丰富度指数最高;而在饥饿培养情况下细菌能利用更多种类的碳源.     

粘细菌的活性检测及与环境因子的相关性

为筛选具有抗菌活性的粘细菌,利用兔粪诱导、大肠杆菌划线诱导和滤纸诱导3种方法从内蒙古凉城地区采集的20种土样中分离粘细菌,结合形态观察、生理生化特征和分子鉴定方法确定这些菌株的分类地位,通过平板对峙法分析抗菌活性,并采用统计学方法分析其与环境因子的相关性.共分离得到140株菌,包括39株纯粘细菌,经鉴定分别为黄色粘球菌(17株)、橙色粘球菌(13株)、变绿粘球菌(5株),还有4株只鉴定到粘球菌属(Myxococcus sp.).抗菌结果显示:39株纯菌均表现出可以对一种甚至多种指示菌产生抗性,其中37株抑制大肠杆菌的生长,35株抑制马铃薯晚疫病菌的生长,26株抑制金黄色葡萄球菌的生长,20株抑制酿酒酵母菌的生长,19株抑制枯草芽孢杆菌的生长.在分析环境因子与粘细菌分布情况的关系时发现,分离出的溶细菌类群的菌株数、溶纤维素类群的菌株数及总的菌株数与土壤p H和速效钾的含量呈正相关,与含水量呈负相关,与其它参数无明显相关性.综上,内蒙古凉城地区粘细菌的优势菌群为粘球菌属,且绝大多数都有不同程度的抗菌活性;结果可为丰富内蒙古地区粘细菌资源及抗马铃薯晚疫病农药研发奠定基础.     

含硫苯衍生物对发光细菌的毒性效应及其定量结构活性相关分析

利用发光细菌法测定了29种氯取代和硝基取代的苯硫(砜、亚砜)基乙酸酯的毒性,通过定量结构活性相关(QSAR)分析发现:此类化合物的毒性主要与其取代基的电荷效应相关,电荷的区域分布对化合物的毒性效应有重要影响.推测该化台物的毒性机理是毒物分子与发光反应中的主要辅酶FMNH_2形成了氢键.还发现:在这类链状毒物分子中,远离活性中心一端的基团具有脱毒作用.     

利用发光细菌测试方法评价菲和芘生物降解过程的毒性变化

将发光细菌毒性测试方法应用于典型多环芳烃生物降解过程的毒性评价,研究了菲和芘单独及混合状态下生物降解过程中的毒性变化。结果表明:溶解态的菲和芘在一定浓度范围内对发光细菌均具有毒性,菲和芘对发光细菌的10 min-EC50值分别为1.61×10-5mol·L-1和2.31×10-6mol·L-1。单组分和混合组分的菲和芘经过微生物降解后,其毒性显著降低。混合组分生物降解体系中由于菲和芘分子之间的互相作用,使其对发光细菌的毒性效应与其单独存在时有差异。生物降解的各阶段,混合组分降解体系的毒性均大于单组分菲或芘降解体系的毒性,且在降解完全的后期,培养液的毒性有增加的趋势。这与菲和芘降解中间产物的积累变化有关。     

Larval settlement in the serpulid polychaete Hydroides elegans in response to bacterial films: an investigation of the nature of putative larval settlement cue

Larval settlement in the marine polychaete Hydroides elegans (Haswell) is induced by certain bacteria in marine biofilms. The exact nature of the settlement cue that larvae of H. elegans receive from these bacteria remains unknown. In this study, we revealed some properties of the bacterially derived larval settlement cue by investigating the larval settlement inductive activity of two bacterial strains after various treatments. These two bacterial strains, Roseobacter sp. and an α-subclass Proteobacteria, are highly inductive to larval settlement of H. elegans. The larvae responded similarly to Roseobacter and Proteobacteria in all the larval settlement bioassays, suggesting that the larval settlement-inducing substances produced by these bacteria may share common characteristics. First of all, the larvae did not settle in the seawater conditioned by the bacteria attached as a film or by the bacteria that were freely suspended in seawater. The results suggest that the putative larval settlement cue is not released into seawater and, therefore, should be associated with the surface of the bacteria. Secondly, formaldehyde treatment entirely eliminated the larval settlement induction activity of the bacterial films, and streptomycin treatment reduced the percentage of larval settlement on the bacterial films in a concentration-dependent manner. Since both treatments can kill bacteria with little damage to the surface chemistry of bacterial cells, the decline in larval settlement is suggested be due to a reduction of the viable bacterial population in the bacterial films. In fact, the reduction of larval settlement in the streptomycin treatments coincided with the decrease in viable bacterial populations in broth cultures containing respective concentrations of streptomycin. These results suggest that the viability of Roseobacter and Proteobacteria is important to their settlement induction effect. Since the larval settlement induction activity of the bacterial strains appears to correlate with their viability, we suggest that the putative larval settlement cue is derived from a metabolic pathway in the bacteria and that the cue is exported to and concentrated at the extracellular polymer matrix of the bacterial cell, at which the larvae establish contact with the bacteria. The larval settlement cue may be highly susceptible to degradation so that a metabolically active bacterial film is needed to maintain the putative cue at a concentration that surpasses the threshold for induction of larval settlement. Received: 14 October 1998 / Accepted: 5 September 2000     

Bacterial communities of the marine sponges Hymeniacidon heliophila and Polymastia janeirensis and their environment in Rio de Janeiro, Brazil

In this study we performed a survey of the bacterial communities associated with the Western Atlantic demosponges Hymeniacidon heliophila and Polymastia janeirensis, based on 16S rRNA sequencing and transmission electron microscopy (TEM). We compared diversity and composition of the sponge-associated bacteria to those of environmental bacteria, represented by free-living bacterioplankton and by bacteria attached to organic particulate matter in superficial sediments. Partial bacterial 16S rRNA sequences from seawater, sediment, and sponges were retrieved by PCR, cloning, and sequencing. Sequences were subjected to rarefaction analyses, phylogenetic tree construction, and LIBSHUFF quantitative statistics to verify coverage and similarity between libraries. Community structure of the free-living bacterioplankton was phylogenetically different from that of the sponge-associated bacterial assemblages. On the other hand, some sediment-attached bacteria were also found in the sponge bacterial community, indicating that sponges may incorporate bacteria together with sediment particles. Rare and few prokaryotic morphotypes were found in TEM analyses of sponge mesohyl matrix of both species. Molecular data indicate that bacterial richness and diversity decreases from bacterioplankton, to particulate organic sediment, and to H. heliophila and P. janeirensis. Sponges from Rio de Janeiro harbor a pool of novel and exclusive sponge-associated bacterial taxa. Sponge-associated bacterial communities are composed of both taxons shared by many sponge groups and by species-specific bacteria.     

Organic and inorganic phosphorus uptake by bacteria in a plug-flow microcosm

Phosphorus (P) is a vital nutrient for sustaining natural water productivity. Both particulate and dissolved forms of organic and inorganic P are potentially important sources of bioavailable P for primary and secondary producers. A microcosm system to imitate the bacterial community in Plym river sediment and pore water is described and bacterial uptake rates for inorganic and organic phosphorus are presented in this paper. The aim of this study was to investigate the uptake of two organic phosphorus compounds (phytic acid and D-glucose-6-phosphate) by freshwater bacteria. The bioreactors comprise glass columns packed with two types of small glass beads on which bacterial biofilm can develop. The glass beads with different porosity were introduced to simulate River SPM. The selected P compounds spiked into the inflow of the microcosm, and measured the step change of P concentration in the outflow to investigate the behavior of bacterial uptake of nutrients. The results showed that organic phosphorus was converted into inorganic phosphorus but the conversion rate depended on the type of phosphorus species. One experiment suggested that phytic acid (refractory) could displace phosphate from the biofilm surface; the other experiment showed that D-glucose-6-phosphate (labile) could be hydrolysed and utilized easily by the bacteria. The results also suggested that bacteria might break down the C-P bonds to utilize the carbon. Further experiments should investigate the effect of varying the C:N:P ratio in the microcosm system to determine which nutrient limits bacteria uptake.     

基于高通量测序的工业园区地下水和土壤细菌群落结构比较研究

为探究工业园区地下水和土壤细菌群落结构、多样性变化特征,采用高通量测序技术对地下水和土壤细菌16S r RNA基因高变区域进行序列测定。通过对Alpha多样性、物种组成、丰度和群落结构的分析,比较地下水和土壤细菌群落结构的异同。Alpha多样性的比较结果表明,土壤细菌群落多样性和丰富度明显高于地下水,地下水细菌群落多样性指数反映出地下水已受到周边污染源的影响。物种注释结果表明,地下水样品共检出48个细菌门,土壤样品共检出50个细菌门。变形菌门(Proteobacteria)、拟杆菌门(Bacteroidetes)和厚壁菌门(Firmicutes)是地下水细菌群落的优势类群,共占93.54%,且该工业园区地下水细菌群落呈现出典型的淡水种群特征;土壤中优势细菌门为Proteobacteria、放线菌门(Actinobacteria)、酸杆菌门(Acidobacteria)、Firmicutes和芽单胞菌门(Gemmatimonadetes),共占85.21%。由于地下水和土壤两者的生态系统和理化环境的差异,致使Actinobacteria、Acidobacteria、绿弯菌门(Chloroflexi)、硝化螺旋菌门(Nitrospirae)、α-变形菌纲(Alphaproteobacteria)、δ-变形菌纲(Deltaproteobacteria)和Gemmatimonadetes占比在地下水和土壤细菌群落间差异显著,同时使地下水和土壤细菌群落各含有一些特有的优势细菌属(地下水2个,土壤4个)。基于高通量测序技术对工业园区样品的测序结果可以为地下水和土壤环境的生态评价提供方法依据。     

细菌偶氮还原研究进展

细菌偶氮还原是在细菌偶氮还原酶作用下偶氮复合物分解为芳香氨的过程,多种细菌都具有偶氮还原功能.有氧条件下细菌的偶氮还原是在特异性的偶氮还原酶作用下完成的,对氧气不敏感.而厌氧菌和兼性厌氧菌的偶氮还原是在厌氧或兼性厌氧条件下由一些氧化还原中间体作为电子穿梭体与偶氮复合物作用的非特性的还原过程.电子穿梭体与偶氮复合物作用是一个纯化学氧化还原过程,其中偶氮复合物作为电子受体接受电子.厌氧偶氮还原是细菌偶氮还原的主要形式.偶氮还原在厌氧-好氧染料废水处理系统和偶氮水溶胶聚合物作为结肠靶向给药载体的研究中有重要应用.图2表1参45     

The microbiological control of mine waste pollution

The removal of heavy metals and neutralisation of acidity in mine waste waters by algae and bacteria are reviewed. Algal growths in artificial meander systems have been shown to remove significant quantities of heavy metals and fine particulates from mine waste waters. Bacterial systems for the oxidation of ferrous iron and the reduction of sulphate leading to neutralisation of acidity are described. The use of bacterial extracellular polymers in the removal of heavy metals from solution is discussed with reference to its application to acid mine waters. It is concluded that a combination of bacterial and algal systems with process optimisation, including nutrient amendment, may provide suitable treatment for mine waste waters. Metal-resistant bacteria may also be important in reclamation, where it is possible that they can confer some degree of tolerance on the plants used to revegetate metal-contaminated tailings dumps.     

Insect chemical communication: Pheromones and exocrine glands of ants

Summary Chemical communication plays a very important role in the lives of many social insects. Several different types of pheromones (species-specific chemical messengers) of ants have been described, particularly those involved in recruitment, recognition, territorial and alarm behaviours. Properties of pheromones include activity in minute quantities (thus requiring sensitive methods for chemical analysis) and specificity (which can have chemotaxonomic uses). Ants produce pheromones in various exocrine glands, such as the Dufour, poison, pygidial and mandibular glands. A wide range of substances have been identified from these glands.     

The effect of conventional wastewater treatment on the levels of antimicrobial-resistant bacteria in effluent: a meta-analysis of current studies

Antimicrobial agents in the environment are a cause for concern. Antimicrobial drug residues and their metabolites reach the aquatic and terrestrial environment primarily through wastewater treatment plants (WWTP). In addition to the potential direct negative health and environmental effects, there is potential for the development of antimicrobial-resistant bacteria. Residue levels below the minimum inhibitory concentration for a bacterial species can be important in selection of resistance. There is uncertainty associated with resistance formation during WWTP processing. A meta-analysis study was carried out to analyse the effect of WWTP processing on the levels of antimicrobial-resistant bacteria within bacterial populations. An analysis of publications relating to multiple antimicrobial-resistant (MAR) bacteria (n?=?61), single antimicrobial-resistant (SAR) E. coli (n?=?81) and quinolone/fluoroquinolone-resistant (FR) bacteria (n?=?19) was carried out. The odds-ratio (OR) of MAR (OR?=?1.60, p?<?0.01), SAR (OR?=?1.33, p?<?0.01) and FR (OR?=?1.19, p?<?0.01) bacteria was determined. The results infer that WWTP processing results in an increase in the proportion of resistant bacteria in effluent, even though the overall bacterial population may have reduced (i.e. a reduction in total bacterial numbers but an increase in the percentage of resistant bacteria). The results support the need for further research into the development of antimicrobial-resistant strains and possible selective pressures operating in WWTPs.     

Characteristics of bacterial community in Poyang Lake sandy land under different vegetation restoration conditions北大核心CSCD

为掌握不同植被恢复措施下鄱阳湖沙地土壤细菌群落变化特征,以湿地松、香根草、单叶蔓荆、狗牙根等植物恢复措施的沙地土壤为研究对象,采用高通量测序技术,分析植物恢复前后细菌群落变化.结果显示,相较对照组,土壤细菌多样性均有提升,在门水平上,绿弯菌门丰度大幅降低,而变形菌门、放线菌门和厚壁菌门丰度增加.在属水平上,副伯克霍尔德氏菌属、慢生根瘤菌属、芽孢杆菌属、分枝杆菌属等丰度提升,而假单胞菌属丰度大幅下降.植被恢复后,不同植被固氮功能细菌丰度提升,而香根草、狗牙根土壤磷细菌丰度下降,湿地松土壤固氮、解磷功能优势细菌提升幅度为76.11%、51.12%,单叶蔓荆为65.04%、23.17%.土壤容重、有机质、全磷、有效氮是影响功能细菌变化的重要因素,其中有效氮与芽孢杆菌属、分枝杆菌属呈显著水平以上相关性.本研究表明鄱阳湖沙地植被恢复措施提升了固氮细菌的丰富度,但不同植被土壤解磷细菌群落变化差异大;研究结果可为沙地治理提供科学依据.(图6表3参34)     

Using T-RFLP technology to analyze bacterial diversity in the rhizospheric soils of tea tree at different ages北大核心CSCD

The rhizospheric soils of Tieguanyin at different ages were used as the study materials, and terminal-restriction fragment length polymorphism (T-RFLP) was used to analyze the changes in bacterial diversity. The results showed that the number of T-RFs, Simpson index, and Shannon index decreased significantly with the age of the tea tree. Results of correlation analysis showed that 9 T-RFs from the bacterial community were significantly and positively correlated with the age of the tea tree, and included 34 species of microbes belonging to 10 classes. The 34 microbes were divided into 6 types according to their functional attributes, and included pathogenic bacteria, bacteria that improved soil texture or inhibited pathogenic bacteria, and bacteria associated with the carbon, nitrogen, or sulfur cycles, in which the percentage of pathogenic bacteria was 58.82%. Seventeen T-RFs were significantly and negatively correlated with the age of the tea tree, and included 38 species of bacteria belonging to 12 classes. The 38 bacterial species were divided into 5 types according to their functional attributes, and included pathogenic bacteria, bacteria that improved soil quality or inhibited pathogenic bacteria, and bacteria associated with the carbon cycle or nitrogen cycle, in which all the others, barring the pathogenic bacteria, accounted for 78.95% of the bacterial population. In brief, the diversity and function of bacteria in the rhizospheric soil of tea tree changed significantly with the age of the tea tree, which provides a theoretical basis for studying the interactions of bacterial communities. © 2018 Science Press. All rights reserved.