中度嗜盐菌Halomonas sp.BYS-1启动子的克隆和测序

提取了中度嗜盐菌Halomonassp.BYS1的基因组DNA,以甲基对硫磷水解酶基因(mpd)为报告基因,以启动子探针pUCmpd为载体,通过鸟枪法在E.coliDH5α中构建了BYS1的启动子文库.通过筛选获得了17个阳性克隆,编号为P1～P17.测定了阳性克隆的甲基对硫磷水解酶(MPH)活性,结果表明,P3中mpd基因的启动子活性最强,它的酶活高达2554.3U/mg,而P17中mpd基因的启动子活性最弱,它的酶活只有68.3U/mg.对P3、P8、P17克隆中的重组质粒的插入片段进行了测序和在线启动子预测.图4表1参17     

无锡太湖新城污水处理厂改良型A^2／O工艺的调试与运行

介绍了无锡市太湖新城污水处理厂的一期工程设计概况和所采用的改良型A^2/O脱氮除磷工艺，分析、探讨了污水处理厂的调试、培菌、试运行过程中出现的问题，并针对性地采用了一系列的解决措施。由监测数据表明，该工艺运行可靠，出水水质稳定，实现了污水处理厂的正常运行。     

活性污泥高质量RNA快速提取方法研究

通过比较RNA产量、纯度、降解程度、特定基因的扩增能力、微生物多样性等评价指标,考察和探讨了5种不同RNA提取方法对活性污泥总RNA提取效果的影响并最终建立了一种快速、有效的活性污泥RNA提取方法,即在TENP和PBS洗涤沉淀污泥的基础上,分别采用溶菌酶和TRIzol裂解活性污泥细菌、氯仿去除细菌裂解液中的蛋白和大部分DNA、异丙醇沉淀核酸和DNase I水解残留DNA后,最后进一步用离心柱纯化RNA.结果表明,这种方法可以有效提取高质量的菌群RNA,不仅提取的RNA总量多(每g污泥可提取169.6μg RNA)、纯度高、降解程度低、完整性好、具有丰富的生物多样性,而且可同时进行16SrRNA和amoA基因的RT-PCR扩增反应;与其它方法相比,性价比高,具有明显的优越性,适用于活性污泥RNA的大量提取,同时,T-RFLP结果证明RNA提取方法对分析样品的微生物种类和丰度分析结果影响较大,不同的RNA提取方法所获得的微生物群落基因多样性及其种类、丰度均不同.本研究建立了一种快速、有效的高质量RNA提取方法,将在监测活性污泥菌群动态变化、菌群代谢功能学、微生物群落芯片等研究上具有重要意义.     

硝化细菌应用于人工湿地生态系统的研究

将亚硝酸细菌和硝酸细菌混合固定于人工湿地系统中,进行了氨氮去除的试验研究。在湿地系统进入稳定运行阶段时,可以观察到系统对于氨氮的去除率稳定在70％左右。同时系统对COD也有着较高的去除率,基本实现了同一系统中的有机物和氨氮的共同去除。     

pUCD-recA重组发光菌构建及对遗传毒性污染物响应作用

基因重组发光菌在水质毒性的评价中具有重要的作用,本研究从分析污染物毒性损伤的机制出发,构建新型pUCD-recA基因重组发光菌.用PCR法从大肠杆菌W3110中扩增recA基因,将其与pGEM-T easy载体连接后测序.测序正确的recA片段及pUCD615载体均用BamHⅠ、EcoRⅠ双酶切,连接后电转化导入宿主菌JM109.挑取克隆,提取质粒用PCR鉴定,阳性克隆再进行测序.将构建成功的pUCD-recA载体转化入大肠杆菌RFM443,加入相应的遗传毒性污染物,观察发光响应作用.结果表明,recA基因PCR扩增出的片段为293 bp,测序结果与GenBank中的recA序列进行BLAST比对,同源性为99%,表明扩增序列正确.与pUCD615载体连接后的测序结果表明,recA基因已正确地插入到pUCD615的多克隆位点,方向和读码框正确,重组发光菌载体构建成功.将构建好的重组载体转化入RFM443宿主菌,加入遗传毒性污染物观察响应效果.丝裂霉素C(MMC)对pUCD-recA重组发光菌诱导效果最好,0.01 mg/L即可有很好的响应曲线;N′-甲基-N′-硝基亚硝基胍(MNNG)则在50~100 mg/...     

一株兼性自养菌硫转化性能的研究

研究了一株具有硫转化性能的兼性自养菌的培养特性,以及不同因素对该菌硫转化纺的影响,结果表明：有机物可促进细菌的生长,合适的氮源、适宜的ｐＨ、添加微量元素,可提高该菌的硫转化率。在约１％接种量时,硫转化率可达８７．４２％。     

固定化微生物菌种的筛选与鉴定

以菲、芘作为降解对象 ,对多种菌株进行筛选 ,从中挑选出降解率最高的两株菌 ,细菌和真菌各一株 ,并对其生长曲线进行测定 ,为菌种的固定化提供了一定的依据。同时对菌种进一步鉴定 ,细菌为动胶杆菌 (Zoogloeasp .) ,在 72h内对菲、芘的降解率为 87 82 %和 4 9 2 2 % ;真菌为镰刀菌 (Fuariumsp .) ,在 2 16h内对菲、芘的降解率为 93 32 %和 96 2 9%。     

Cellular damage of plant pathogenic fungi by antifungal compounds produced by Bacillus spp. isolates

The use of microorganisms as biological control agents (BCAs) has become an effective alternative to chemical means of controlling plant pathogens. The antagonistic and inhibitory activity of 71 Bacillus spp. strains, which were isolated from different Mexican sites, were tested against several phytopathogen fungi, Fusarium oxysporum, Fusarium equiseti, Fusarium avenaceum, Bipolaris spp. and Alternaria spp. From the antagonism study, the strain ELI149 showed a marked inhibition of growth against all tested fungi; therefore crude metabolites from this strain were extracted using ethyl acetate and amberlite resin and probed against the same fungi as well as strains of Mucor sp., Penicillium spp. and Paecilomyces spp. The results indicated that amberlite was more suitable for extraction of secondary metabolites with antifungal activity. Finally, observation of cell damage in the tested pathogenic fungi showed marked morphological changes on reproductive structures in all tested fungi indicating that antibiosis was the mechanism of the antagonistic effect. These results suggest that metabolites from the Bacillus strains have a wide spectrum of antibiotic activities, which can be used as biocontrol agents for controlling fungal plant diseases of agricultural importance.     

Changes of microbial composition during wastewater reclamation and distribution systems revealed by high-throughput sequencing analyses

This study employed 454-pyrosequencing to investigate microbial and pathogenic communities in two wastewater reclamation and distribution systems. A total of 11972 effective 16S rRNA sequences were acquired from these two reclamation systems, and then designated to relevant taxonomic ranks by using RDP classifier. The Chao index and Shannon diversity index showed that the diversities of microbial communities decreased along wastewater reclamation processes. Proteobacteria was the most dominant phylum in reclaimed water after disinfection, which accounted for 83% and 88% in two systems, respectively. Human opportunistic pathogens, including Clostridium, Escherichia, Shigella, Pseudomonas and Mycobacterium, were selected and enriched by disinfection processes. The total chlorine and nutrients (TOC, NH₃-N and NO₃-N) significantly affected the microbial and pathogenic communities during reclaimed water storage and distribution processes. Our results indicated that the disinfectant-resistant pathogens should be controlled in reclaimed water, since the increases in relative abundances of pathogenic bacteria after disinfection implicate the potential public health associated with reclaimed water.     

促进氰钴胺素转化为腺苷钴胺素的基因表达与鉴定

辅酶B12作为甘油脱水酶(GDHt)的辅酶参与到Klebsiella pneumoniae代谢甘油生成1,3-丙二醇(1,3-PD)和3-羟基丙酸(3-HP)的途径中.前期研究表明底物甘油可以导致辅酶B12分子中Co—C键的断裂,进而引起GDHt的失活.为进一步研究非活性的维生素B12(CNCbI)转化为具有活性的辅酶B12(AdoCbI)的过程,从K.pneumoniae中克隆得到了ATP:钴(I)胺素腺苷转移酶(ACA)基因btuR和还原酶基因yciK,并成功构建了双启动子表达质粒pUC18-tac-btuR-tac-yciK,将表达质粒转化Escherichia coli JM109获得了一株重组菌.利用改进的分析方法,结果表明:1)重组蛋白具有腺苷转移酶的活性;2)重组菌可以很好地将非活性的钴胺素,如维生素B12,转化生成具有活性的辅酶B12.