家蚕丝心蛋白轻链cDNA和基因调控片段的克隆及序列分析

从家蚕品种菁松×皓月后部丝腺中提取总RNA ,用RT -PCR扩增技术克隆到家蚕丝心蛋白轻链cDNA .序列分析表明 ,该片段全长为 798个碱基 .比较 4种不同品种来源的家蚕丝心蛋白轻链基因外显子区域序列 ,同源性在98.0 %～ 99.4 %之间 ,推测的氨基酸同源性在 98.0 %～ 99.6 %之间 ,4个品种间发生的变异比较一致地集中在不同位置的 8个碱基上 .提取家蚕品种菁松×皓月后部丝腺总DNA ,进行PCR反应 ,获得家蚕丝心蛋白轻链基因调控片段 ,长度约为 1.2kb,测定了其邻近起始密码子一端包括 5 76个碱基的DNA序列 .结果表明 ,该片段包括一些典型的调控元件和多个可能参与丝蛋白基因特异性表达调控的元件 ,该部分序列与J - 139L链基因相应区域同源性高达 98.8% .图 3参 8     

粘红酵母(Rhodotorula glutinis CIBAS A 1401)苯丙氨酸解氨酶cDNA核心序列的克隆与分析

L苯丙氨酸因其在工业、医药上的重要作用而日益受到人们的重视.苯丙氨酸解氨酶(PAL)生物转化肉桂酸生产L苯丙氨酸成了该领域研究与开发的热点.本文利用简并寡核苷酸PCR法结合cDNA末端快速扩增PCR法,从粘红酵母中克隆了苯丙氨酸解氨酶cDNA核心序列,为克隆其cDNA全长序列及体外表达其活性蛋白奠定了基础.图5参22     

L-山梨糖还原酶基因在大肠杆菌中的表达及融合蛋白的酶学特性

为获得L 山梨糖还原酶 (SR)整个操纵子序列 ,以 pGEM 3zf( )作为载体构建了Gluconobactersp .S6基因文库 .结合转化子在以L 山梨糖为唯一碳源培养基上的生长情况 ,利用PCR技术筛选到一个含L 山梨糖还原酶基因的阳性克隆pGEM 3zf( ) sr3/E .coli,酶切鉴定插入片断长度约 5 .0kb左右 .以 pGEM 3zf( ) sr3/E .coli质粒DNA为模板 ,扩增SR结构基因 .PCR产物经测序验证为SR基因序列后 ,与表达载体Pet32a( )相连 ,转化宿主大肠杆菌AD4 94 (DE3)对SR基因进行表达 ,并利用重金属离子亲和层析技术对SR融合蛋白进行了纯化 ,对其酶学特性进行了研究 .结果表明 :还原型辅酶II(NADPH)是SR酶蛋白的最适电子供体 ;SR酶蛋白的最适反应pH为 7.0 ,pH为6 .5时保持稳定 ,酶活力较高 ;最适反应温度是 5 0℃ ,30℃时热稳定性较好 .SDS PAGE电泳分析 ,SR融合蛋白的Mr 在6 5× 10 3 左右 ,由此推测出天然SR的Mr 在 5 3× 10 3 左右 ,与SR基因序列推测出的分子量大小一致 .图 13表 2参 6     

Start-up of the anammox process from the conventional activated sludge in a hybrid bioreactor

The anaerobic ammonium oxidation (anammox) process was successfully started up from conventional activated sludge using a hybrid bioreactor within 2 months. The average removal efficiencies of ammonia and nitrite were both over 80%, and the maximum total nitrogen removal rate of 1.85 kg N/(m³.day) was obtained on day 362 with the initial sludge concentration of 0.7 g mixed liquor suspended solids (MLSS)/L. Scanning electron microscope (SEM) observation of the granular sludge in the hybrid reactor clearly showed a high degree of compactness and cell sphericity, and the cell size was quite uniform. Transmission electron microscope photos showed that cells were round or oval, the cellular diameter was 0.6--1.0 μupm, and the percentage of the anammoxosome compartment was 51%--85% of the whole cell volume. Fluorescence in situ hybridization analysis (FISH) indicated that anammox bacteria became the dominant population in the community (accounting for more than 51% of total bacteria on day 250). Seven planctomycete 16S rRNA gene sequences were present in the 16S rRNA gene clone library generated from the biomass and affiliated to Candidatus Kuenenia stuttgartiensis and Candidatus Brocadia sp., a new anammox species. In addition, the average effluent suspended solid (MLSS) concentrations of outlets I (above the non-woven carrier) and II (below the non-woven carrier) were 0.0009 and 0.0035 g/L, respectively. This showed that the non-woven carrier could catch the biomass effectively, which increased biomass and improved the nitrogen removal rate in the reactor.     

Effects of nonylphenol on brain gene expression profiles in F1 generation rats

The effects of nonylphenol (NP) were examined on brain gene expression profiles of F1 generation rats by means of microarray techniques. The mRNA were extracted from the brain of two-day-old F1 generation rats whose F0 male generation were treated with NP, then reversely transcribed to cDNA and labeled with Cy5 and Cy3 fluorescence. Subsequently, the cDNA probes were hybridized to the mouse 40S cDNA microarray and the fluorescent signals of Cy5 and Cy3 were scanned and analyzed. Sixteen identified genes were expressed significantly differently from control, including 13 down-regulated, of which five were related to brain energy metabolism. Data suggest that it is possible that NP affects function of energy metabolism in male rats when administered perinatally.     

Building spectral libraries for wetlands land cover classification and hyperspectral remote sensing

Recent advances in remote sensing provide opportunities to map plant species and vegetation within wetlands at management relevant scales and resolutions. Hyperspectral imagers, currently available on airborne platforms, provide increased spectral resolution over existing space-based sensors that can document detailed information on the distribution of vegetation community types, and sometimes species. Development of spectral libraries of wetland species is a key component needed to facilitate advanced analytical techniques to monitor wetlands. Canopy and leaf spectra at five sites in California, Texas, and Mississippi were sampled to create a common spectral library for mapping wetlands from remotely sensed data. An extensive library of spectra (n=1336) for coastal wetland communities, across a range of bioclimatic, edaphic, and disturbance conditions were measured. The wetland spectral libraries were used to classify and delineate vegetation at a separate location, the Pacheco Creek wetland in the Sacramento Delta, California, using a PROBE-1 airborne hyperspectral data set (5m pixel resolution, 128 bands). This study discusses sampling and collection methodologies for building libraries, and illustrates the potential of advanced sensors to map wetland composition. The importance of developing comprehensive wetland spectral libraries, across diverse ecosystems is highlighted. In tandem with improved analytical tools these libraries provide a physical basis for interpretation that is less subject to conditions of specific data sets. To facilitate a global approach to the application of hyperspectral imagers to mapping wetlands, we suggest that criteria for and compilation of wetland spectral libraries should proceed today in anticipation of the wider availability and eventual space-based deployment of advanced hyperspectral high spatial resolution sensors.     

Stoichiometric and molecular evidence for the enrichment of anaerobic ammonium oxidizing bacteria from wastewater treatment plant sludge samples

Anammox enrichments were readily developed from seven municipal wastewater treatment plants (WWTPs) sludge, but not with methanogenic granular sludge from two agro-industrial WWTPs. Only 50 d was required for the first evidence of anammox activity from a return activated sludge obtained from a WWTP operated for nutrient removal. The molar ratios of nitrite and ammonium consumption of approximately 1.32 as well as nitrate and dinitrogen gas product ratios of approximately 0.095 provided evidence of the anammox reaction. The presence of anammox was confirmed by polymerase chain reaction (PCR) using primer sets (PLA46F and AMX820R) specific for anammox bacteria. The 16S rRNA gene fragment of anammox bacteria was detected in seven enrichment cultures (ECs) with demonstrated anammox activity but not in the original inocula from which the ECs were derived and also not in the two methanogenic sludge samples, which indicates the PCR predicted the anammox activity. Two genera, Brocadia and Kuenenia, were successfully identified as the Planctomycetes occurring in the clone libraries of successful anammox enrichments. Brocadia dominated in cultures that were respiked extensively; whereas Kuenenia predominated in cultures that were less aggressively respiked. These findings indicate that respiking management may play an important role on selecting the genus of anammox bacteria. The batch enrichment results clearly illustrate that anammox can be readily enriched from municipal sludge from a wide variety of process operations at WWTPs.     

浅议图书馆对继续教育的服务

继续教育是我院教育教学工作的重要组成部分。继续教育生具有"自学为主、限时多学、积极多研、追求速成",但利用图书馆较少的特点。图书馆应结合实际,以提高对继续教育的服务质量为宗旨,从转变思想观念,加强制度建设和馆藏建设,以及如何进行图书宣传等方面为继续教育服务。     

黑曲霉(ASPERGILLUS NIGER)SG株三磷酸甘油醛脱氢酶基因的分离与初步鉴定

黑曲霉（Ａｓｐｅｒｇｉｌｕｓｎｉｇｅｒ）在工业生产和科学研究中占有重要地位,因为许多黑曲霉菌株能产生有机酸和酶制剂,并能分泌大量蛋白质到细胞外［１,６］．三磷酸甘油醛脱氢酶（Ｇｌｙｃｅｒａｌｄｅｈｙｄｅ３ｐｈｏｓｐｈａｔｅｄｅｈｙｄｒｏｇｅｎａ...     

同时产甲烷反硝化颗粒污泥中微生物群落结构

在UASB成功运行同时产甲烷反硝化小试基础上,针对反应器内颗粒污泥,通过构建古菌和细菌的16SrDNA基因文库、RADAR遗传变异分型和测序比对等技术进行了微生物系统发育关系和群落结构分析.结果表明,在颗粒污泥的古菌中,产甲烷髦毛菌和产甲烷杆菌是主要菌群,随机选出的88个古菌克隆子,这两类古菌的16SrDNA序列分别占古菌总量71.59%和22.73%;而颗粒污泥中细菌的多样性要高于古菌,低GC革兰氏阳性菌和ε变型菌纲分支的细菌是细菌的主要菌群,在随机选出的133个细菌克隆子中,这2类细菌的16SrDNA序列分别占细菌总量的49.62%和12.03%.