Classification of subcellular location by comparative proteomic analysis of native and density-shifted lysosomes

One approach to the functional characterization of the lysosome lies in the use of proteomic methods to identify proteins in subcellular fractions enriched for this organelle. However, distinguishing between true lysosomal residents and proteins from other cofractionating organelles is challenging. To this end, we implemented a quantitative mass spectrometry approach based on the selective decrease in the buoyant density of liver lysosomes that occurs when animals are treated with Triton-WR1339. Liver lysosome-enriched preparations from control and treated rats were fractionated by isopycnic sucrose density gradient centrifugation. Tryptic peptides derived from gradient fractions were reacted with isobaric tag for relative and absolute quantitation eight-plex labeling reagents and analyzed by two-dimensional liquid chromatography matrix-assisted laser desorption ionization time-of-flight MS. Reporter ion intensities were used to generate relative protein distribution profiles across both types of gradients. A distribution index was calculated for each identified protein and used to determine a probability of lysosomal residence by quadratic discriminant analysis. This analysis suggests that several proteins assigned to the lysosome in other proteomics studies are not true lysosomal residents. Conversely, results support lysosomal residency for other proteins that are either not or only tentatively assigned to this location. The density shift for two proteins, Cu/Zn superoxide dismutase and ATP-binding cassette subfamily B (MDR/TAP) member 6, was corroborated by quantitative Western blotting. Additional balance sheet analyses on differential centrifugation fractions revealed that Cu/Zn superoxide dismutase is predominantly cytosolic with a secondary lysosomal localization whereas ATP-binding cassette subfamily B (MDR/TAP) member 6 is predominantly lysosomal. These results establish a quantitative mass spectrometric/subcellular fractionation approach for identification of lysosomal proteins and underscore the necessity of balance sheet analysis for localization studies.     

广州市十种森林生态系统的碳循环

为了探讨南亚热带森林生态系统碳循环的规律,在广泛收集资料和试验数据的基础上,对广州10种森林生态系统的碳循环进行研究.结果表明：10种森林生态系统的碳密度在108.35～151.85 t C·hm^-2,其中乔木层碳密度在10.85～48.86 t C·hm^-2,0～60 cm土壤层在87.74～99.01 t C·hm^-2,均低于全国平均水平;从大气流向植被层的碳流量为4.41～9.15 t C·hm^-2·a^-1,植被层流向土壤层的碳流量为0.74～2.06 t C·hm^-2·a^-1,土壤层流向大气层的碳流量为3.94～5.42 t C·hm^-2·a^-1,即系统从大气净吸收碳在0.47～4.97 t C·hm^-2·a^-1之间.各种林分的净系统生产力不同,阔叶林大于针叶林,混交林大于纯林,天然次生林大于人工林.     

Strong repulsive forces between protein and oligo (ethylene glycol) self-assembled monolayers: a molecular simulation study

Restrained molecular dynamics simulations were performed to study the interaction forces of a protein with the self-assembled monolayers (SAMs) of S(CH2)4(EG)4OH, S(CH2)11OH, and S(CH2)11CH3 in the presence of water molecules. The force-distance curves were calculated by fixing the center of mass of the protein at several separation distances from the SAM surface. Simulation results show that the relative strength of repulsive force acting on the protein is in the decreasing order of OEG-SAMs > OH-SAMs > CH3-SAMs. The force contributions from SAMs and water molecules, the structural and dynamic behavior of hydration water, and the flexibility and conformation state of SAMs were also examined to study how water structure at the interface and SAM flexibility affect the forces exerted on the protein. Results show that a tightly bound water layer adjacent to the OEG-SAMs is mainly responsible for the large repulsive hydration force.     

植物抗虫基因工程研究进展

植物抗虫基因工程为防治农业害虫提供了一条崭新途径。本文对植物抗虫基因工程近年来所取得的某些研究进展,包括目前已发现和利用的抗虫基因、提高抗虫基因在植物体内表达的方法以及防止或延缓害虫产生抗性的策略等方面进行了综合评述,并对植物抗虫基因工程中有待解决的问题和发展前提提出了自己的看法。     

HCV E2蛋白诱导的体液免疫及CTL应答研究

应用PCR方法扩增出HCVE2基因编码417a.a～750a.a的DNA片段,克隆到原核表达载体pQE30 LacZ启动子下游,转化JM109菌株.在JM109菌株中诱导表达出N端含6个组氨酸的E2融合蛋白,用Ni-NTA-Superflow亲和层析柱纯化作为抗原免疫实验兔和BALB/c鼠.定期取免血,采用间接ELISA方法检测兔子体内针对E2的抗体水平和维持规律.结果显示,距初次免疫14d兔子体内已有抗体产生,直至免疫第55d抗体水平持续上升,之后抗体水平保持稳定,抗体滴度达到13200.六周后,取鼠脾脏制备淋巴细胞,定向刺激扩增后与经过重组真核表达质粒pCE2转染的P815细胞作用,利用LDH释放试验检测作用效果.在ET=2001的情况下,杀伤率超过30%.这些结果表明工程菌株表达的HCV E2蛋白具有良好的免疫原性,可以诱发免疫实验动物机体产生较高滴度的抗体及特异性CTL应答.由此我们认为E2蛋白是发展HCV预防工程蛋白疫苗的合适候选者.     

Genetic map of Triticum turgidum based on a hexaploid wheat population without genetic recombination for D genome

ABSTRACT: BACKGROUND: A synthetic doubled-haploid hexaploid wheat population, SynDH1, derived from the spontaneous chromosome doubling of triploid F1 hybrid plants obtained from the cross of hybrids Triticum turgidum ssp. durum line Langdon (LDN) and ssp. turgidum line AS313, with Aegilops tauschii ssp. tauschii accession AS60, was previously constructed. SynDH1 is a tetraploidization-hexaploid doubled haploid (DH) population because it contains recombinant A and B chromosomes from two different T. turgidum genotypes, while all the D chromosomes from Ae. tauschii are homogenous across the whole population. This paper reports the construction of a genetic map using this population. RESULTS: Of the 606 markers used to assemble the genetic map, 588 (97%) were assigned to linkage groups. These included 513 Diversity Arrays Technology (DArT) markers, 72 simple sequence repeat (SSR), one insertion site-based polymorphism (ISBP), and two high-molecular-weight glutenin subunit (HMW-GS) markers. These markers were assigned to the 14 chromosomes, covering 2048.79 cM, with a mean distance of 3.48 cM between adjacent markers. This map showed good coverage of the A and B genome chromosomes, apart from 3A, 5A, 6A, and 4B. Compared with previously reported maps, most shared markers showed highly consistent orders. This map was successfully used to identify five quantitative trait loci (QTL), including two for spikelet number on chromosomes 7A and 5B, two for spike length on 7A and 3B, and one for 1000-grain weight on 4B. However, differences in crossability QTL between the two T. turgidum parents may explain the segregation distortion regions on chromosomes 1A, 3B, and 6B. CONCLUSIONS: A genetic map of T. turgidum including 588 markers was constructed using a synthetic doubled haploid (SynDH) hexaploid wheat population. Five QTLs for three agronomic traits were identified from this population. However, more markers are needed to increase the density and resolution of this map in the future study.     

哑蟋属两新种及峨眉哑蟋雌虫的发现(直翅目：蟋蟀科)

哑蟋属两新种及峨眉哑蟋雌虫的发现（直翅目：蟋蟀科）陈军,郑哲民（陕西师范大学动物研究所西安７１００６２）１９９３年在整理陕西师范大学动物研究所蝗虫标本室的蟋蟀标本以及作者于１９９２年夏季在四川采集的蟋蟀标本时,发现有哑蟋属ＧｏｎｉｏｇｒｙｌｌｕｓＣｈ...     

广州市农作物系统与大气的CO2交换

在广泛收集资料和实验分析的基础上,研究了广州市各种农作物系统与大气CO2交换.分析了各种农作物系统净生产力吸收CO2的能力和碳汇功能大小.结果表明:2005年广州市8种农作物系统作物净生产力吸收CO2 4 032 366t·a-1,其土壤CO2排放3981753t·a-1,吸收大于排放,对大气CO2而言,整个农作物系统是一个弱的碳汇;水稻、甘蔗、木薯和果用瓜4种连作或高杆作物系统每年作物净生产力吸收CO2量大于土壤CO2的排放量,系统具有较大的碳汇功能,花生、大豆、花卉和蔬菜4种矮杆作物系统每年作物净生产力吸收CO2量小于土壤CO2的排放量,系统起着碳源作用;果实或经济产量生长在地上部分的作物其单位面积吸收CO2能力比果实(块根)生长在地下的作物大;除花生在生育期间生物量吸收CO2量少于同期土壤排放以外,其余7种作物在生育期间生物量吸收CO2的量大于同期土壤排放,大多数农作物在生育期间具有碳汇功能,在撂荒期才体现碳源作用.     

山楂蜂花粉中脂肪酸的GC-MS分析

用乙醇-氯仿作溶剂,提取出山楂蜂花粉粗脂肪,然后进行甲酯化处理,用气相色谱/质谱联用技术进行分离鉴定,共鉴定出10种脂肪酸,并测定相对含量,多不饱和脂肪酸含量较高,其中亚油酸相对含量为47.84%,亚麻酸相对含量为19.30%.