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1.
采用廉价的乳糖替代IPTG诱导重组别藻蓝蛋白(rAPC)在大肠杆菌(Escherichiacoli)JM109中表达,对诱导产物表达的培养基、培养条件、诱导剂的浓度和诱导时机进行了优化,将优化条件用于5L自控发酵罐进行高密度培养。结果表明,对rAPC表达的条件进行优化后,乳糖诱导rAPC的表达率可达26.8%,与IPTG的诱导结果接近;在高密度培养中,OD600最大达21.8。研究结果可为乳糖替代IPTG大规模生产rAPC奠定基础。 相似文献
2.
Hueiwang Anna C. Jeng Andrew J. Englande Reda M. Bakeer Henry B. Bradford 《Estuarine, Coastal and Shelf Science》2005,63(4):513-526
Urban stormwater runoff could contribute to the deterioration of water quality of a receiving water body. In this research, field studies and laboratory experiments were conducted to assess the microbial contamination resulting from urban stormwater runoff into the Lake Pontchartrain estuary. Fecal coliform, Escherichia coli and enterococci were used as indicator organisms. The specific objectives of the research were to examine the distribution of the indicator organisms in different environmental elements (water column, suspended particles and sediment) and to further investigate the mechanisms related to their fate. Results of the research indicated satisfactory water quality at the study sites during dry weather periods. However, a significant increase was observed in the concentrations of the indicator organisms in the water columns and sediment at specific study sites following a given stormwater event. Three to seven days were needed for the elevated indicator organisms to return back to their background levels in the water column and sediment, respectively. The mechanism of sedimentation contributed to a reduction in the microbial concentration in the water column, as the indicator organisms were found to attach to the suspended particles in the stormwater. The percentage of fecal coliform, E. coli and enterococci attached to the suspended particles was found to be within the range of 9.8–27.5%, 21.8–30.4%, and 8.4–11.5% of the total indicator organisms in the stormwater loaded into the estuary, respectively. About 75–80% of the total indicator organisms remained free-floating for some distance in the water column before dying off. 相似文献
3.
乳糖诱导重组别藻蓝蛋白基因在大肠杆菌中的表达 总被引：1，自引：0，他引：1
以重组别藻蓝蛋白表达工程菌JM109（DE3）／pET28-APC作为研究对象，对于JM109（DE3）菌株在乳糖诱导下表达重组蛋白的规律进行了研究。比较分析了不同生长阶段进行诱导，最佳乳糖浓度、诱导持续时间和诱导温度等参数对重组蛋白表达的影响。实验结果表明，采用JM109（DE3）为宿主菌，经过条件优化，乳糖诱导重组蛋白的表达量可以达到IPTG诱导的水平；较低的诱导培养温度能有效提高可溶性重组蛋白的表达。最后在摇瓶发酵结果的基础上，实现了乳糖诱导工程菌的5L全自动发酵罐培养。研究结果为在大肠杆菌大规模发酵中，运用廉价、无毒的乳糖作为诱导剂提供了一定的借鉴。 相似文献
4.
溶藻弧菌asp基因在大肠杆菌中表达活性研究及条件优化 总被引：1，自引：0，他引：1
为进一步研究溶藻弧菌碱性丝氨酸蛋白酶(ASP)蛋白生物学活性和功能,将构建的pET23d-asp在大肠杆菌BL21(DE3)中表达,对表达产物进行纯化分析,并优化表达条件。结果表明:在咪唑洗脱缓冲液浓度为100 mmol/L时,表达的可溶性ASP被大量洗脱,纯化的ASP相对分子质量约为42 000,具有蛋白活性;在诱导温度28℃、异丙基-β-D硫代半乳糖苷(Isopropyl-β-D-thiogalactopyranoside,IPTG)浓度1 mmol/L及诱导时间16 h时,表达的活性ASP蛋白最多。 相似文献
5.
通过分析海绵清除大肠杆菌的过程,研究海绵净化细菌的机理。作者利用荧光显微镜和激光共聚焦显微镜观测等手段,监测和分析了绿色荧光大肠杆菌(Escherichia coli)在繁茂膜海绵(Hymeniacidon perlevis)体内、体外水环境中数量变化过程。在1 L含有3×107个/m L绿色荧光大肠杆菌的海水中放入鲜重(1.02±0.11)g的繁茂膜海绵24块,处理7 h,海水中的荧光大肠杆菌数量逐渐降低;而海绵体内荧光大肠杆菌数量在2 h时内逐渐增多,之后的2 h趋于稳定,4 h以后开始逐渐减少。水体中大肠杆菌不仅进入海绵体内,而且进入海绵细胞内。含有荧光大肠杆菌的海绵块转入无菌海水中后,海绵体内及细胞中大肠杆菌逐渐消失,而且大肠杆菌没有被释放到环境海水中。分析表明,繁茂膜海绵能够以摄食的方式净化水环境中的大肠杆菌。 相似文献
6.
Bonilla TD Nowosielski K Cuvelier M Hartz A Green M Esiobu N McCorquodale DS Fleisher JM Rogerson A 《Marine pollution bulletin》2007,54(9):1472-1482
Fecal indicator levels in nearshore waters of South Florida are routinely monitored to assess microbial contamination at recreational beaches. However, samples of sand from the surf zone and upper beach are not monitored which is surprising since sand may accumulate and harbor fecal-derived organisms. This study examined the prevalence of fecal indicator organisms in tidally-affected beach sand and in upper beach sand and compared these counts to levels in the water. Since indicator organisms were statistically elevated in sand relative to water, the study also considered the potential health risks associated with beach use and exposure to sand. Fecal coliforms, Escherichia coli, enterococci, somatic coliphages, and F(+)-specific coliphages were enumerated from sand and water at three South Florida beaches (Ft. Lauderdale Beach, Hollywood Beach, and Hobie Beach) over a 2-year period. Bacteria were consistently more concentrated in 100g samples of beach sand (2-23 fold in wet sand and 30-460 fold in dry sand) compared to 100ml samples of water. Somatic coliphages were commonly recovered from both sand and water while F(+)-specific coliphages were less commonly detected. Seeding experiments revealed that a single specimen of gull feces significantly influenced enterococci levels in some 3.1m(2) of beach sand. Examination of beach sand on a micro-spatial scale demonstrated that the variation in enterococci density over short distances was considerable. Results of multiple linear regression analysis showed that the physical and chemical parameters monitored in this study could only minimally account for the variation observed in indicator densities. A pilot epidemiological study was conducted to examine whether the length of exposure to beach water and sand could be correlated with health risk. Logistic regression analysis results provided preliminary evidence that time spent in the wet sand and time spent in the water were associated with a dose-dependent increase in gastrointestinal illness. 相似文献
7.
We previously described the genomic structure of the cytochrome P450 1A (CYP1A) gene from the hermaphroditic fish Rivulus marmoratus [Kim, I.-C., Kim, Y.J., Yoon, Y.-D, Kawamura, S., Lee, Y.-S., Lee, J.-S., 2004a. Cloning of cytochrome P450 1A (CYP1A) genes from the hermaphroditic fish Rivulus marmoratus and the Japanese medaka Oryzias latipes. Mar. Environ. Res. 58, 125–129]. To further characterize R. marmoratus CYP1A, we cloned the cDNA sequence of a CYP1A gene from this species and also expressed its recombinant protein in an E. coli system. We exposed R. marmoratus to 4-nonylphenol, and found a small induction of CYP1A mRNA in the treated animals. In this paper, we discuss the characteristics of R. marmoratus CYP1A gene as well as its potential use in a biomonitoring assay. 相似文献
8.
Recent studies have suggested that poikilothermic animals, such as fish, may represent a previously overlooked source of the faecal indicator bacteria (FIB) used for the assessment of water quality. However, quantitative studies of FIB in poikilotherms are scarce. We investigated the presence of FIB in the faeces of five freshwater fish species. E. coli and enterococci were detected in 71 and 76% of faecal samples, respectively. Concentrations were highly variable, with maximum concentrations (2.1?×?104 E. coli and 1.3?×?106 enterococci per gram of faecal material) up to four orders of magnitude higher than present in the water column. FIB were not detected in faecal samples from marine fish. Our findings suggest that FIB ingested from the environment may be capable of replication within the fish gut, making fish a potential source and transport mechanism for FIB in New Zealand freshwaters. This may have implications for water quality monitoring. 相似文献
9.
不同种类纳米零价铁的毒性比较研究 总被引：1，自引：1，他引：0
纳米零价铁是一种高效的环境修复材料,可以处理多种污染物;然而,纳米粒子的尺寸效应可能导致其在自然界中存在潜在毒性风险。选择几种常用包覆型、负载型和裸露的纳米铁,通过大肠杆菌的耐受性实验,比较3种纳米铁的毒性。研究表明,负载型纳米铁的分散性最好,而裸露纳米铁最差。3种纳米铁虽然对大肠杆菌都表现出毒性,但是负载型纳米铁的毒性最小。通过毒性减缓的机理分析,说明纳米铁改性后阻止了纳米颗粒与细菌的直接接触,这是空间位阻效应的作用。研究结果进一步证实了在使用纳米材料前应充分评估潜在毒性和环境效应的重要性。 相似文献
10.
采用Ames试验分别对蒙脱石、石英、方解石、钠长石4种可吸入矿物细颗粒及其与大肠杆菌、表皮葡萄球菌相互作用48 h后的混悬液作为受试物进行检测,分析IMG是否具有致突变性,及其与人体正常细菌群作用后,IMG/正常菌复合体是否有致突变性毒力改变。结果显示石英以及石英/大肠杆菌复合体对TA98、TA100菌株Ames试验呈阳性反应;石英/表皮葡萄球菌复合体对TA98菌株Ames试验呈阳性反应;而蒙脱石、方解石、钠长石纯矿及其与正常菌作用后的复合体Ames试验结果均为阴性。结果表明石英具有直接致突变作用;石英与正常菌菌作用后复合体依然具有致突变性,但石英表现出的碱基置换突变作用在与表皮葡萄球菌作用后消失,其机制有待进一步研究。 相似文献