苏云金芽孢杆菌和大肠杆菌穿梭质粒的构建和特性研究

将苏云金芽孢杆菌中的pHTA1030质粒与大肠杆菌中的pJH101质粒重组后,构建出pBHGA重组质粒。此质粒通过逐步酶切,缺失后重组得到了14个分子量大小不同的衍生重组质粒。经过对pBHG1重组穿梭质粒在E.coli HB101和B.subttlis 168受体中表达的分析,证明了它带有B.thuringiensis(简写作B.t.)质粒的启动区、启始复制区和对热分离稳定区基因片段,并能高频转化B.t.受体细胞和高表达外源cat基因,同时具有对热分离稳定的特性。为B.t.基因工程体系提供了高效转化表达载体。     

一株抗锰土壤芽胞杆菌的分离鉴定与生物学活性分析

从山东崅屿采集的黄棕壤中分离得到一株具有抗Mn(Ⅱ)和Mn(Ⅱ)氧化双重活性的芽胞杆菌,其最高Mn(Ⅱ)耐受浓度达到130mmol/L,对Mn(Ⅱ)的氧化活性为3.3μmol/(L·d)。通过个体形态与培养特征观测、生理生化反应、G+Cmol%测定和16SrDNA序列比对分析等鉴定,确定该菌株为巨大芽胞杆菌(Bacillus megaterium),命名为MB283。该菌株在添加Mn(Ⅱ)(10mmol/L)条件下比不添加Mn(Ⅱ)表现出相对较快的生长速率。采用高温培养并结合0.01%SDS处理,从MB283菌株筛选到一株发生内生质粒消除的突变株MB287,具有与野生菌株类似的锰耐受活性,且对Mn(Ⅱ)的氧化活性与野生菌株相比无明显改变,表明野生菌株MB283中与锰抗性和锰氧化相关的基因可能是定位于该菌的染色体上。     

多基因SNP位点及多种危险因素与老年汉族冠心病的关联分析

目的:探索老年冠心病多基因遗传易感性基础及相关的危险因素。方法:采用病例-对照研究方法,共入选老年汉族冠心病患者246例,非冠心病患者185例,纳入性别、年龄、吸烟,饮酒,高血压史、糖尿病史、高脂血症史、同型半胱氨酸、氨基末端脑钠肽前体、超敏C反应蛋白、抗凝血酶III、胆固醇、甘油三酯、高密度脂蛋白、低密度脂蛋白共15种危险因素与冠心病的关联性进行logistic回归分析。同时使用美国Sequenom高通量基因多态性分型技术研究了10种基因11个单核苷酸基因多态性(SNP)位点与冠心病的关联性。结果:15种危险因素中,发现增龄、高血压、抗凝血酶III(ATIII)下降是冠心病主要的危险因素,P<0.05。11个SNPs中3个SNP,血小板糖蛋白GP1BA rs2243093(-5T/C),血管紧张素转化酶ACE rs4332(547C/T)与ATIII rs2227589(893C/T)与老年汉族患者冠心病相关联。rs2243093(-5T/C)突变基因型CC与TT+AT比较,P=0.029(OR=3.41,CI:1.19-9.75);rs4332(547C/T)杂合型TC与CC+TT相比,P=0.003(OR=0.56,CI:0.38-0.82);rs2227589(893C/T),CT+CT与野生基因型CC相比较,P=0.003(OR=1.79,CI:1.22-2.63)。结论:增龄、抗凝血酶III下降、高血压是影响老年冠心病的主要危险因素,血小板、抗凝血系统、肾素-血管紧张素系统三种机制参与了老年冠心病的发生与发展。     

荒漠植物白刺属4个物种的生殖分配比较

选定乌兰布和沙漠地区白刺属4种植物为研究对象,通过对其样株在花期的各生殖构件的数量特征及生物量调查,系统研究了唐古特白刺(Nitraria tangutorum Bobr.)、西伯利亚白刺(Nitraria sibirica Pall.)、大白刺(Nitraria roborowskii Kom.)和泡泡刺(Nitraria sphaerocarpa Maxim.)4种白刺属植物在生殖枝水平上的生殖分配。结果表明:不同白刺属植物在分株高度、生殖枝长、生殖枝基径、单枝花数、花序干重、枝叶干重等生殖构件的数量性状方面均有显著差异,其中泡泡刺的各生殖构件的数量均最小;除了西伯利亚白刺的生殖分配值达到44.51%外,其余3种白刺的生殖分配值均没有超过20%。经统计分析,4种白刺种群的生殖枝长分别与分株高度呈显著(P<0.05)的直线性正相关关系;生殖枝花序干重与分株高显著正相关;4种白刺的生殖分配随着分株生殖枝生物量的增加而减少,即白刺的个体大小与生殖分配之间呈现负相关关系。这种生殖分配特点反映了不同白刺植物对生长环境的资源利用、与克隆繁殖的权衡及对生态适应的策略。     

自絮凝酵母高浓度重复批次乙醇发酵

利用发酵性能优良的自絮凝酵母Saccharomyces cerevisiaeflo,研究开发了重复批次高浓度乙醇发酵系统,以节省下游加工过程的能耗。在终点乙醇浓度达到120g/L左右的条件下,发酵系统的乙醇生产强度达到8.2g/(L·h)。然而实验中发现,随着发酵批次的增多,自絮凝酵母沉降性能逐渐下降,从发酵液中沉降分离所需时间相应延长,导致发酵液中高浓度乙醇对酵母的毒害作用加剧,影响其发酵活性和发酵系统运行的稳定性,发酵装置运行11个批次后无法继续运行。实验结果表明,絮凝能力下降导致的酵母絮凝颗粒尺度减小是其沉降性能下降的主要原因。进一步研究发现,酵母的絮凝能力通过再培养可以恢复。在此基础上对发酵系统操作进行改进,每批发酵结束后可控采出一定比例菌体,调节系统的酵母细胞密度和乙醇生产强度以刺激酵母增殖,保持其絮凝能力。在达到相同发酵终点乙醇浓度条件下,虽然发酵系统的乙醇生产强度降低到4.0g/(L·h),但运行10d后絮凝颗粒酵母尺度趋于稳定,继续运行14d,未发现絮凝颗粒酵母尺度继续下降的现象,系统可以稳定运行。     

Development of mechanistically based model for simulating soluble microbial products generation in an aerated/non-aerated SBR

Soluble microbial products (SMPs) are considered as the main organic components in wastewater treatment plant effluent from biological wastewater treatment systems. To investigate and explore SMP metabolism pathway for further treatment and control, two innovative mechanistically based activated sludge models were developed by extension of activated sludge model no.3 (ASM3). One was the model by combining SMP formation and degradation (ASM3-SMP model) processes with ASM3, and the other by combining both SMP and simultaneous substrate storage and growth (SSSG) mechanisms with ASM3 (SSSG-ASM3-SMP model). The detailed schematic modification and process supplements were introduced for comprehensively understanding all the mechanisms involved in the activated sludge process. The evaluations of these two models were demonstrated by a laboratory-scale sequencing batch reactor (SBR) operated under aerated/non-aerated conditions. The simulated and measured results indicated that SMP comprised about 83% of total soluble chemical oxygen demand (SCOD) in which biomass-associated products (BAPs) were predominant compared with utilization-associated products (UAPs). It also elucidated that there should be a minimum SMP value as the reactive time increases continuously and this conclusion could be used to optimize effluent SCOD in activated sludge processes. The comparative results among ASM3, ASM3-SMP and SSSG-ASM3-SMP models and the experimental measurements (SCOD, ammonia and nitrate nitrogen) showed clearly the best agreement with SSSG-ASM3-SMP simulation values (R = 0.993), strongly suggesting that both SMP formation and degradation and SSSG mechanisms are necessary in biologically activated sludge modeling for municipal wastewater treatment.     

Genetic and alkaloid analysis of Menispermum dauricum DC. by RAPD and HPLC

The aim of the present work was to determine whether dauricine could be used as a taxonomic marker for Menispermum dauricum DC., and to explore the correlation among RAPD, ecological markers and chemical markers. To this end, the chemical and genetic differences of 173 individual samples of M. dauricum from nine different sources were studied based on the relevant ecological factors including longitude, latitude, annual precipitation, mean temperature, annual accumulated temperature and mean sea level. The contents of dauricine in the sample rhizomes were assayed by HPLC with photodiode array detection. The leaves from the same sample were assayed using randomly amplified polymorphism DNA (RAPD). The genetic distances were then compared. Hierarchical cluster analysis and multiple linear stepwise regression analysis were used in the statistical analysis. The results indicated that the contents of dauricine were respectively correlated with the genetic distance (r = 1.000), longitude (r = 0.849), latitude (r = 0.861), annual precipitation (r = 0.903), mean temperature(r = 0.912), annual accumulated temperature (r = 0.919) and mean sea level (r = 0.925). It is concluded that the content of dauricine in M. dauricum is significantly correlated with genetic distance and ecological factors, and may be used as the taxonomic marker.     

饲养方式对异色瓢虫幼虫生存的影响

在实验室条件下,用不同饲养器具和隔离物来饲养异色瓢虫并对其幼虫生长发育进行了比较。结果表明：低龄幼虫饲养器具以密闭型为好,可减少水分的蒸发;高龄幼虫的饲养器具以透气型为好,有利于幼虫后期的生长发育。饲养密度以每盒（16cm&#215;12cm&#215;6cm）30头为宜。饲养阻隔物以纸扇为佳,可促进高龄幼虫化蛹,缩短幼虫的发育历期。     

Overexpression of phosphoenolpyruvate carboxylase from Jatropha curcas increases fatty acid accumulation in Nicotiana tabacum

Jatropha curcas L. is an excellent biofuel crop, which displays a high efficiency of carbon absorption, and seed oil of Jatropha can be efficiently processed to produce high-quality biodiesel. Plant phosphoenolpyruvate carboxylases (PEPCs) play important roles not only in initial fixation of atmospheric CO₂ in C₄ and Crassulacean acid metabolism (CAM) plants, but also in fatty acid biosynthesis in seeds of oil plants by regulating carbon partitioning. Here, we identified JcPEPC1 from J. curcas L. by homology cloning, and alignment analysis of protein sequence revealed JcPEPC1 was a plant C₃-type PEPC, and shared high similarity to PEPC of castor oil plant Ricinus communis. We implemented detailed functional characterization of JcPEPC1 by expression analysis and transgenic tobacco. JcPEPC1 gene expressed in the leaves and seeds of J. curcas L., and remarkable increase of expression level was also detected at seed oil-accumulating stages. We overexpressed JcPEPC1 in tobacco, and showed the enzymatic activity of PEPC in transgenic plants was notably higher than wild type. Gas chromatography (GC) analysis elucidated the composition and total content of fatty acids were also altered. This study indicated JcPEPC1 played a fundamental role in fatty acid biosynthesis in Jatropha seeds. Our results proposed enhanced PEPC activity of Jatropha could improve biosynthesis of fatty acid, which implied critical functions in primary metabolism of non-photosynthetic PEPC.