Studies on the binary and ternary complexes formed by a neurospora glutamate dehydrogenase and its substrates

The NADP⁺ specific glutamate dehydrogenase from wild-type $\text{Neurospora crassa}$ forms a stable binary complex with NADPH. This can combine with L-glutamate, α-ketoglutarate or the substrate analogue D-glutamate to form ternary complexes which can be distinguished by their different fluorescence properties. The affinity of the enzyme for NADPH diminishes with increases in pH or ionic strength of the solution. Experimental data obtained using modified glutamate dehydrogenases from mutant strains of $\text{N. crassa}$ suggest that the reduced-coenzyme binding sites observed fluorimetrically are the same as those observed by enzyme kinetics.     

水葫芦根际细菌群落结构多样性分析

【目的】了解水葫芦根际细菌群落结构。【方法】运用末端限制性片段长度多态性(Terminal restriction fragment length polymorphism,T-RFLP)技术分析富营养化水体中水葫芦根际和水葫芦近、远水样的细菌群落特征及多样性,结合克隆文库技术和培养法分析根际的细菌种群类型。【结果】同一时期水葫芦根际细菌多样性(Shannon-Weiner指数H′或Simpson指数D)更高,水葫芦近水样次之,远水样最小。10月份的细菌多样性高于5月份的。通过水葫芦根际细菌的克隆文库可知变形杆菌门(Proteobacteria)是水葫芦根际细菌的主要类群,占总群体的65.1%,包括噬菌弧菌(Bacteriovorax sp.)、Dechloromonas sp.、Leptothrix sp.、红螺菌科(Rhodospirillaceae)、Rhodoferax sp.和红环菌科(Rhodocyclaceae)等。T-RFLP图谱显示159 bp为最大优势菌,247 bp为第二大优势菌,对照克隆文库及培养结果分析247 bp属于γ-Proteobacteria,159 bp为不动杆菌(Acinetobacter sp.)。【结论】水葫芦根际细菌的群落结构丰富,不同时段水葫芦根际细菌的丰度略有变化,主要类群为变形杆菌门。     

维生素E对萼花臂尾轮虫繁殖的影响

15℃下 ,添加 Ve浓度为 2 0～ 80 ng/ m l时 ,种群密度与培养时间呈显著相关 ,其中 2 0 ng/ ml和 4 0 ng/ ml组最大种群密度分别为 76 .4 0 ind/ ml和 6 5 .2 0 ind/ ml,显著高于对照组 (最大密度 12 .0 0 ind/ ml)。最大混交雌体百分率低于 10 % ,对照组最大混交雌体比率为 4 0 %。轮虫卵雌比有不同程度的提高 ,但维生素浓度为 10 0 ng/ ml时 ,对种群增长不利 ;2 5℃下 ,对照组和各添加组最大密度依次为 6 8.0 0 ,12 2 .0 0 ,12 4 .0 0 ,16 8.0 0和 2 36 .0 0 ind/ ml。最大卵雌比依次为 1.2 1,1.2 5 ,1.2 2 ,1.4 1和 0 .6 9,混交雌体百分比分别为 2 7.6 9,19.89,18.4 2 ,17.0 2和 12 .88% ;30℃下 ,最大密度依次为 113.80 ,12 1.0 0 ,12 2 .80和 15 1.2 0 ind/ ml,最大卵雌比依次为 1.5 0 ,1.4 2 ,0 .6 4和 0 .96。混交雌体百分比分别为 1.76 ,32 .2 6 ,37.5 0和 32 .0 0 %。     

荒漠植物准噶尔无叶豆结实、结籽格局及其生态适应意义

连续2a采用野外记录的方式对荒漠植物准噶尔无叶豆的结实、结籽格局进行了比较研究。结果表明:植株内花序生成格局表现为不同大小的花序在总花序中的比重与其结实率存在显著正相关,其中含有中等花数目的花序所占的比重及其结实率均最高,此种分布格局能够最大程度的保证繁殖成功;果序内果实生成格局表现为单花着生位置(从近柄端算起)与其结实量占总结实量的比例成线性负相关,花序基部的结实比例最高,顶部结实比例近为0;荚果内种子生成格局表现为:中间位置的胚珠败育率最低。另外,年际间荚果内仅有(1.08±0.03)粒和(1.07±0.03)粒种子能够完全成熟,此种结籽格局是准噶尔无叶豆保证后代质量的最佳策略。     

沙枣（Elaeagnus angustifolia）和孩儿拳头（Grewia.biloba G.Don var.parviflora）幼苗气体交换特征与保护酶对干旱胁迫的响应

以沙枣和孩儿拳头2年生盆栽苗为材料,采用称重控水的方法设置4个土壤含水量梯度(CK、T1、T2、T3),研究不同干旱胁迫对沙枣和孩儿拳头气体交换特征与保护酶的影响.结果显示:(1)干旱胁迫不仅引起两物种净光合速率、蒸腾速率、气孔导度、胞间 CO2 浓度的下降,而且使其日变化曲线在一定程度上发生改变;在轻度(T1)和中度胁迫(T2)下,两物种净光合速率下降主要是由气孔因素引起的,重度胁迫(T3)下,净光合速率下降主要是非气孔因素引起的.(2)随着干旱胁迫增加,沙枣瞬时水分利用效率呈现增加下降再增加趋势,孩儿拳头呈现下降趋势;两物种表观光能利用效率显著下降,重度胁迫下(T3),下降率达50%左右,孩儿拳头表观光能利用效率对干旱胁迫比较敏感;两物种表观CO2利用效率总体呈现下降趋势,沙枣表观CO2利用效率日进程经历了单峰(T1)、双峰(T2)、单峰(T3)的变化,孩儿拳头各处理的表观CO2利用效率日变化均呈现单峰曲线.(3)随着干旱胁迫加剧,两物种叶片的超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT)活性先升高后降低,土壤含水量高于12.8%时,两物种SOD酶活性均高于CK,随着土壤含水量的降低,SOD酶活性低于CK;重度胁迫下(T3),沙枣POD酶活性虽然有所下降,但仍高于CK,而孩儿拳头则和CK无显著差异;两物种CAT酶活性在重度胁迫下(T3)显著低于CK;随着干旱胁迫程度的增加,两物种叶片中的丙二醛(MDA)含量均呈现升高趋势,孩儿拳头脂质过氧化程度受干旱胁迫的影响较大.     

姜黄素通过抑制凋亡信号调节激酶1减弱PM2.5短期暴露致大鼠子宫损伤

细颗粒物(PM2.5)是空气动力学直径≤2.5 μm的颗粒物,能诱发多种疾病.已有大量的流行病学调查证实,PM2.5能够损伤生殖系统,但其致病机制不明确,相关的研究也非常有限.为研究PM2.5短期暴露对大鼠子宫的损伤,以及姜黄素(curcumin,CRC)对其保护作用,本研究将50只雌性SD大鼠随机分为生理盐水对照组、...     

四种鹭类繁殖生态生物学研究

本文对夜鹭,白鹭,黄鹭白鹭,池鹭混群在同一巢区的繁殖,种间协高及雏鸟生长进行进行了观察研究,其中黄嘴白路豫南大别山类新纪录,在此繁殖亦为首次发现。几种路在巢位,食性,取食活动和习性方面都表现出其混居同一巢地繁殖的种间协调性,符合高期假说。比较４种鹭鸟雏鸟生长过程中各生长参数。其中池鹭虽个体最小,但体重和各部位生长却对较快。     

幽门螺杆菌尿素酶B亚单位基因工程菌发酵工艺的研究

通过不同培养基、不同葡萄糖浓度、不同溶氧条件、补料与非补料对幽门螺杆菌尿素酶B亚单位(UreB)基因工程菌的菌体生长与外源蛋白表达量的影响的比较 ,建立了稳定、适宜的幽门螺杆菌尿素酶B亚单位基因工程菌发酵工艺。多批实验结果证明 ,菌体单产可达 86 g/L ,目的蛋白的表达率为 38.2 %。     

Structural variation in human apolipoprotein E3 and E4: secondary structure, tertiary structure, and size distribution

Human apolipoprotein E (apoE) is a 299-amino-acid protein with a molecular weight of 34 kDa. The difference between the apoE3 and apoE4 isoforms is a single residue substitution involving a Cys-Arg replacement at residue 112. ApoE4 is positively associated with atherosclerosis and late-onset and sporadic Alzheimer's disease (AD). ApoE4 and its C-terminal truncated fragments have been found in the senile plaques and neurofibrillary tangles in the brain of AD patients. However, detail structural information regarding isoform and domain interaction remains poorly understood. We prepared full-length, N-, and C-terminal truncated apoE3 and apoE4 proteins and studied their structural variation. Sedimentation velocity and continuous size distribution analysis using analytical ultracentrifugation revealed apoE3(72-299) as consisting of a major species with a sedimentation coefficient of 5.9. ApoE4(72-299) showed a wider and more complicated species distribution. Both apoE3 and E4 N-terminal domain (1-191) existed with monomers as the major component together with some tetramer. The oligomerization and aggregation of apoE protein increased when the C-terminal domain (192-271) was incorporated. The structural influence of the C-terminal domain on apoE is to assist self-association with no significant isoform preference. Circular dichroism and fluorescence studies demonstrated that apoE4(72-299) possessed a more alpha-helical structure with more hydrophobic residue exposure. The structural variation of the N-terminal truncated apoE3 and apoE4 protein provides useful information that helps to explain the greater aggregation of the apoE4 isoform and thus has implication for the involvement of apoE4 in AD.