argE基因在大肠杆菌中的表达优化

N-乙酰鸟氨酸脱酰基酶可在重组菌BL21（DE3）-pET22b-argE中表达。首先确定了该酶的细胞表达定位,再研究了诱导温度、诱导剂种类及浓度、诱导起始菌体密度、诱导时间等因素对重组菌生长及目的蛋白表达活性的影响。结果表明,IPTG和乳糖皆可诱导目的蛋白表达,乳糖的诱导效果优于IPTG。在诱导起始0D600为0．46时加入15g／L乳糖,20℃诱导18h最适于目的蛋白的活性表达。表达条件优化后,酶活从1．68U／mL提高至282．99U／mL,约为原来的168倍。     

乳糖诱导耐热木聚糖酶基因在大肠杆菌中表达

考察乳糖代替IPTG诱导耐热木聚糖酶基因在重组大肠杆菌BL21中表达的可行性。分别以IPTG和乳糖作为诱导剂,对诱导时机、诱导剂浓度、诱导持续时间等主要因素进行了分析比较。结果表明,当菌体生长至发酵液吸光值OD600达1．3时加入乳糖其终浓度达29．2 mmol／L,37℃,持续诱导9．5h,木聚糖酶活力达到最高,为3587.5U,是IPTG优化条件下持续诱导7h达到的最高酶活的2．07倍,而成本只有IPTG的1／204。     

小桐子的组织培养和植株再生

以小桐子（Jatropha curcas）的胚芽、子叶、下胚轴、叶柄、叶片和茎段作为外植体,用不同浓度的6-苄基腺嘌呤（6-BA）和α-萘乙酸（NAA）对其进行愈伤组织的诱导和植株再生的研究。结果表明：在MS培养基中加入5．0mg／L6-BA和1．0mg／LNAA对愈伤组织的诱导效果最好;加入5．0mg／L6-BA和0．1mg／LNAA对不定芽的诱导最为有效,加入0．1mg／L6-BA和1．0mg／LNAA有利于芽的生长;加入1．0mg／LNAA的1／2MS培养基对生根最为有利。     

噬菌体T7溶菌酶工程菌的构建

将噬菌体T7溶菌酶基因克隆到含有可诱导性启动子LacUV 5的表达载体pARl206中。在没有诱导剂存在情况下,LacUV 5启动子的转录受到抑制。当带有重组质粒的转化菌生长到适当浓度,向培养液中加入0．4mmo1／L IPTG,LacUV 5启动子受到诱导,从而使插入的基因高水平表达,并积累大量具有生物活性的溶菌酶。噬菌体T 7溶菌酶在大肠杆菌中表达的产量平均约22mg／L。     

安菜的组织培养与植株再生

通过对安菜组织培养的培养基种类、激素配比、碳源的研究,得出适于安菜侧芽分化生长的最佳培养基为MS＋6-BA0．5mg／L＋NAA0．2mg／L。诱导愈伤组织以2,4-D浓度在0．2mg／L～0．5mg／L之间最佳。碳源以蔗糖,浓度3％最佳。以IBA0．5mg／L～1．5mg／L浓度诱导生根效果最好。     

重组戊型肝炎病毒衣壳蛋白工程菌的高密度培养

在10L发酵罐中对戊型肝炎病毒衣壳蛋白在重组大肠杆菌中表达发酵工艺进行了研究,用分批培养方法探讨了不同培养基、培养基中磷酸盐浓度和Mg^2＋浓度等因素对菌体生长与重组蛋白表达的影响;用分批补料培养研究了不同的补料工艺对菌体生长与重组蛋白表达的影响,同时对重组菌诱导时期、诱导持续时间以及不同诱导温度表达包含体在尿素溶液中的溶解性进行了研究。结果表明,在优化后的培养基中,磷酸盐浓度、Mg^2＋浓度分别为80mmol/L 与20mmol/L时菌体生长与表达效果较好;分批补料培养中,37℃培养9h菌体达到对数期中期(约45OD600)为适宜诱导时期,加入终浓度为10mmol/L IPTG后诱导5h,OD600达到80以上,重组蛋白表达量达到29.74％,为最适收获菌体时间;37℃表达的包含体80％以上溶解在4mol/L的尿素溶液中,最终浓度达到14mg/mL; 10L发酵罐中确定的发酵工艺参数在30L发酵罐中进行了放大培养,10L发酵罐中确定的发酵工艺参数在30L发酵罐上具有可放大性与重复性, 可以应用于工业生产。     

脱色希瓦氏菌S12的铁还原性能研究*

从印染废水中分离得到了一株具有染料脱色功能的希瓦氏菌脱色新种。该菌能在厌氧条件下利用Fe3+作为末端电子受体获得能量,支持细胞生长。在pH8．0,温度30℃,柠檬酸铁800mg／L,乳酸钠2g／L,酵母抽提物0．5g／L的条件下,培养8h的过程中,菌体细胞量的增长完全与Fe3+的还原发展趋向一致。同时考察了碳氮源、乳酸钠、酵母抽提物、pH值和温度等方面对该菌株的生长和铁还原特性的影响。结果表明,菌体生长以LB为最好,以葡萄糖和乳酸钠为碳源时对铁还原有利。在酵母抽提物浓度4g／L范围内,菌体生长量和铁还原率     

HCMV pp65截短蛋白原核表达条件优化

为了提高HCMVpp65蛋白片段在大肠杆菌中表达量,研究了不同发酵条件对其表达的影响,包括培养基、接种量、温度、摇床转速、pH、诱导时间以及诱导剂IPTG使用浓度等。结果表明,以LB培养基为发酵液,按5%接种量,37℃培养3h后IPTG诱导5h,重组菌菌体生物量为0.6g/L,目的蛋白表达量达18mg/L。用3.7L发酵罐进行放大培养,菌体生物量达0.85g/L,最高目的蛋白表达量达到25mg/L。     

金属离子对黄柄曲霉生长和抗真菌抗生素合成的影响

不同的金属离子对兼性海洋霉菌黄柄曲霉179的生长有不同的影响。在0．002mol/L的浓度下,Hg^2 、Ag^ 和Cr^3 能强烈抑制该菌的生长,Pb^2 、Sr^2 、Co^2 、Al^3 对其生长有一定的抑制,生长量低于对照;Mn^2 、Ba^2 、Zn^2 对其生长没有明显影响;Cu^2 对该菌的生长有微弱的促进作用。菌体生物量略高于对照;不同的金属离子对曲霉179真菌抗生素179M合成有不同的影响。与对照相比,Ba^2 对共产量没有影响;Al^3 、Zn^2 、Sr^2 有一定的抑制作用,其发酵相对效价分别为对照的81．4％、55．5％和65％;0．002mol／L斩Mn^2 和Pb^2 能强烈抑制此抗生素的合成,在添加0．002mol/LMn^2 和Pb^2 的培养基中,虽然菌体生长良好,但无179M产生;Co^2 和Cu^2 则有明显的促进作用,当培养基中添加0．002ml／L的Co^2 时其发酵相对效价提高到261．4％;当培养基中添加0．003mol／L的Cu^2 时其发酵相对效价提高到350．2％。Co62 和Cu62 对发酵的促进效应是相互拮抗的。     

6种抗真菌药物对皮肤癣菌体外抗真菌活性评价

目的评价6种抗真菌药物对皮肤癣菌体外抗真菌活性。方法采用CLSI推荐的M-38P方案对分离自足癣和体、股癣的皮肤癣菌进行联苯苄唑、硝酸舍他康唑、硝酸异康唑、盐酸布替萘芬、阿莫洛芬、利拉萘酯6种抗真菌药物敏感性测定。结果联苯苄唑MIC范围为0．03—4mg／L,MIC50为1mg／L,MIC90为2mg／L。硝酸舍他康唑分别为0．06—16mg／L、0．5mg／L和2mg／L。硝酸异康唑分别为0．03～2mg／L、0．25mg／L和0．5mg／L。盐酸布替萘芬分别为0．0025～0．04mg／L、0．01mg／L和0．02mg／L。阿莫罗芬分别为0．01～〉0．08mg／L、0．02mg／L和0．04mg／L。利拉萘酯分别为0．004—0．625mg／L、0．039mg／L和0．312mg／L。结论6种抗真菌药物对皮肤癣菌均有强的抗菌活性,由强到弱依次为布替萘芬、阿莫罗芬、利拉萘酯和咪唑类药物。     

Enumeration and presumptive identification of some functional groups of bacteria in the rumen of dairy cows fed grass silage-based diets

Abstract Samples of rumen ingesta from two rumen-fistulated dairy cows fed grass silage-based diets were examined for numbers and types of bacteria that developed colonies on rumen fluid-agar media designated to support the growth of (a) a wide range of species, (b) cellulolytic bacteria, (c) lactate-fermenting bacteria, (d) non-fermentative bacteria. The most numerous species was Bacteroides ruminicola followed by Butyrivibrio fibrisolvens . The most abundant cellulolytic species were Eubacterium cellulosolvens and Ruminococcus flavefaciens. Megasphaera elsdenii and Selenomonas ruminantium were important lactate fermenters but an unidentified bacterium that grew poorly on maintenance medium was by far the most numerous among bacteria isolated from lactate-containing medium. One strain remained sufficiently viable to show that it fermented lactate to propionate and acetate.     

解钾菌与解磷菌及固氮菌的相互作用

微生物是土壤肥力的核心 ,土壤中的微生物不仅数量巨大 ,而且种类极多 ,许多微生物对土壤Ｎ ,Ｐ和Ｋ等养分的转化和供给起非常重要的作用。目前 ,研究比较多的主要有解钾菌、解磷菌和固氮菌。能够使土壤中矿物态钾转化为有效形态的细菌 ,目前发现的主要有扭脱芽胞杆菌 (Ｂａｃｉｌｌｕｓｅｘ ｔｏｒｑｕｅｎｓ)、胶质芽胞杆菌 (Ｂ .ｍｕｃｉｌｇｉｎｏｓｎｓ)、环状芽胞杆菌 (Ｂ .ｃｉｒｃｕｌａｎｓ) [1,2 ] 等 ,这些细菌有些同时能使矿物态的Ｐ转化为有效形态。能够使土壤中无效态磷转化为有效态磷的微生物种类也很多 ,研究比较多的是假单…     

海洋滩涂沉积物环境中几类主要细菌的动态分布

目的研究宁波市宁海县贝类养殖滩涂中异养细菌、反硝化细菌、氨化细菌和硫酸还原菌等几类细菌的动态分布及异养细菌的菌群组成。方法异养细菌的计数采用平板菌落计数法;反硝化细菌、氨化细菌和硫酸还原菌的计数采用MPN法;并参照Oliver提供的海洋细菌鉴定检索图、《海洋调查规范》和《一般细菌常用鉴定方法》提供的图式将分离到的细菌鉴定至属。结果滩涂沉积物环境中异养细菌的数量波动在8.00×103~7.60×104CFU/g(湿重),反硝化细菌的数量变动在5.00×102~5.00×104个/g(湿重),氨化细菌的数量变动在3.00×105~5.00×107个/g(湿重),硫酸还原菌的数量分布在9.00×104~9.00×106个/g(湿重),温度的变化对这几类细菌的数量分布没有直接的影响。所分离的细菌经过鉴定,可归于15个属与肠杆菌科的部分属,其中肠杆菌科(Enterobacteriaceae)的部分属、梭状芽胞杆菌属(Clostridium)、芽胞杆菌属(Bacillus)、棒状杆菌属(Corynebacterium)、发光杆菌属(Photobacterium)、假单胞菌属(Pseudomonas)等为优势菌属。沉积物中氨化细菌、反硝化细菌和硫酸还原菌的检出率均为100%,且含量很高。结论经过多年不间歇的养殖后,宁海贝类养殖滩涂已经不同程度地受到了污染,应尽快调整养殖容量并对养殖环境进行重新布局。     

Mini-review of the concentration variations found inthe alfresco atmospheric bacterial populations

Variations in the atmospheric surface layer's culturable, and to a lesser extent, total bacteria-associated atmospheric particlecharacteristics will be discussed in terms of (a)their temporal variation from 2 min resolutionthrough diurnal to annual periods, (b) the effect ofmeteorological conditions on their abundance andsize, (c) total to culturable bacterial ratio, and(d) the total number of bacteria per culturableparticle (e) bacterial survival in droplet/particles, and (f) the general particle size distribution including aerodynamic Count MedianDiameter (CMD).Meteorological and topographic conditions thatcontrol total and culturable bacteria-associatedatmospheric particle concentration will be presentedin terms of (a) precipitation, (b) wind direction,(c) time of day, (d) sky conditions (i.e., cloudy,sunny, rain, etc.), (e) season, and (f) atmosphericinversion conditions.Simulation models will be described that supporthypotheses of diurnal and annual concentrationcycles in the Earth's (and perhaps other planetaryatmospheres) atmospheric surface layer.     

Aerobic and anaerobic microbial consumption of hydrogen in geothermal spring water

Abstract Hydrogen consumption was measured in hot geothermal water from two ponds of the San Federigo solfatara, Tuscany, Italy, where emanation gases contained approx. 4% H₂. H₂ consumption was completely inhibited by NaOH and partially by HgCl₂ indicating microbial utilization. Aerobic and anaerobic H₂ consumption activities coexisted in the same water with aerobic activity being higher in one pond and anaerobic activity in the other. The kinetics of H₂ consumption were consistent with those of 'Knallgas', methanogenic or sulfidogenic bacteria.     

Distribution of bacteria in the sediment of an oxbow tropical lake (Lagoa do Infernão,SP, Brazil)

The temporal distribution of bacteria on the surface sediment of the oxbow lake Lagoa do Infernão was studied over a period of one year using different techniques for bacterial counts. Several bacterial groups were differentiated, such a total bacteria and filamentous, heterotrophic, amylolytic, and lipolytic bacteria. The best defined temporal distribution pattern was obtained by total-AO bacterial counts, with a variation of 1.26 × 10¹⁰ to 8.58 × 10¹⁰ bacteria (g dw)^–1 during the dry and rainy seasons, respectively. The electrode potential (Eh) of the surface sediment ranged from – 148 mV during the rainy season to + 371 mV during the dry season. The variations detected on the surface sediment suggest a greater allochthonous contribution of material and bacteria resulting in high decomposing activity during the rainy season.     

微生物对油田污水-聚丙烯酰胺体系粘度的影响

比较了厌氧和曝气处理后的油田采出水配制聚丙烯酰胺溶液(采出水-聚丙烯酰胺体系)粘度及该体系中腐生菌,铁细菌和硫酸盐还原菌的数量变化。结果表明,曝气有利于聚合物溶液粘度的保持,粘度损失率明显低于厌氧采出水-聚合物体系。总体上,腐生菌或其代谢产物对聚合物粘度的影响很小;铁细菌可以利用聚合物生长,是破坏聚合物粘度的主要微生物类群;硫酸盐还原菌不能直接利用大分子量聚合物,可以利用小分子或分子链断裂的聚合物生长。     

Characterization of Communities of Heterotrophic Bacteria Associated with Healthy and Diseased Corals in Nha Trang Bay (Vietnam)

A comparative investigation of the heterotrophic microflora of 11 species of healthy corals and of white-band-diseased and yellow-band-diseased corals inhabiting the reefs of Nha Trang Bay (Vietnam), which has been exposed to anthropogenic impact, was performed. Fifty-nine strains of heterotrophic bacteria isolated on Y/K and Endo media were investigated and characterized. All the isolates were identified at the genus level by consideration of the results of analysis of their phenotypic properties, determination of the molar percent of G+C bases in their DNA, and the composition of the fatty acids of lipids. In the composition of the microflora of tissues of healthy corals, γ-proteobacteria prevailed, with halomonads being dominant among them. In addition, the gram-negative bacteria included Pseudomonas and Vibrio spp., members of the Cyto-phaga-Fla-vobacterium-Bacteroides (CFB) phylogenetic cluster, and Moraxella sp. The gram-positive bacteria revealed included Bacillus, Staphylococcus, Halococcus, and Micrococcus spp., and coryneform bacteria. In the composition of the microflora of the tissues of affected corals, bacteria of the family Enterobacteriaceae and of the genera Planococcus and Arthrobacter, which were not revealed in healthy hydrobionts, were found. The anthropogenic impact is not the sole factor determining the infection of corals.     

Proteomics‐based identification of orchid-associated bacteria colonizing the Epipactis albensis,E. helleborine and E. purpurata (Orchidaceae,Neottieae)

Using proteomics-based identification by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS), we conducted the first analysis of the composition of endophytic bacteria isolated from different parts of selected Epipactis species, i.e. the buds, the inflorescences and the central part of the shoots, as well as the rhizomes. We identified aerobic and anaerobic bacteria, including such taxa as Bacillus spp., Clostridium spp., Pseudomonas spp. and Stenotrophomonas spp., which may be considered as promoting plant growth. Because most of the indicated bacteria genera belong to spore-producing taxa (spores allow bacterial symbionts to survive adverse conditions), we suggest that these bacteria species contribute to the adaptation of orchids to the environment. We found clear differences in the microbiome between investigated closely related taxa, i.e., Epipactis albensis, E. helleborine, E. purpurata and E. purpurata f. chlorophylla. Some of the analysed orchid species, i.e. E. albensis and E. purpurata co-occur in habitats, and their bacterial microbiomes differ from each other.