Genomic cloning, expression and recombinant protein purification of alpha and beta subunits of the allophycocyanin gene (apc) from the cyanobacterium Anacystis nidulans UTEX 625

A genomic fragment encoding alpha(APC) and beta(APC) (i.e., alpha and beta units of the allophycocyanin, APC) from Anacystis nidulans UTEX 625 was cloned and sequenced. This fragment, containing a non-coding sequence of 56 nucleotides in between, was then subcloned into the expression vector pMal-c2 downstream from and in frame with the malE gene of E. coli encoding MBP ( maltose binding protein). The fusion protein was purified by amylose affinity chromatography and cleaved by coagulation factor Xa. alpha(APC) and beta(APC) were then separated from MBP and MBP fusion proteins, respectively, and concentrated by membrane centrifugation. The study provides a method to produce recombinant allophycocyanin subunits for biomedical and biotechnological applications.

栉孔扇贝抗鳗弧菌感染性状候选基因的多态性研究

栉孔扇贝（Chlamys farreri）是我国北方地区主要的养殖贝类之一，曾为沿海各省带来巨大的经济效益。但自1997年以来，陆续爆发的病害问题给扇贝养殖业造成了巨大的经济损失，严重影响了该产业的健康发展。目前认为培育抗病性强的扇贝优良品种是解决病害问题的根本途径。由于传统的育种方法费时费力，无法满足对良种的迫切需求，因此有必要通过分子手段加快抗病品种的培育步伐。标记辅助育种（marker assisted selection，MAS）是成功应用于动物育种中的分子手段之一，但由于缺乏与抗病性状相关的标记，MAS目前还无法在软体动物中得到应用。因此，寻找与抗病性状相关的分子标记是在软体动物中发展MAS的关键。 本研究利用鳗弧菌（Listonella anguillarum）对栉孔扇贝进行攻毒感染实验，初步得到敏感群体和抗病群体后采用PCR、PCR-RFLP、Bi-PASA PCR等方法研究了CfLysG、CfC1qDC和CfLITAF基因多态性及其与栉孔扇贝对鳗弧菌抗性的关系。 研究发现，栉孔扇贝CfLysG的基因序列中共有104个单核苷酸多态性（SNP）位点和29个插入/缺失（I/D）多态性位点。有17个多态性位点位于启动子区域，选择其中的-753 I/D、-391A/G和-284I/D多态性进行检测，发现这三个位点的基因型在敏感群体和抗病群体中的分布均符合Hardy-Weinberg平衡（P>0.05）。其中-753 ID基因型和-284 ID基因在抗病群体中的频率高于在敏感群体中的频率，但两者之间无显著性差异（P>0.05）。-391 AG基因型在抗病群体中的频率显著高于敏感群体（P=0.007），表明-391 AG基因型与栉孔扇贝对鳗弧菌的抗性显著相关。为验证这一相关性，对-391位点不同基因型的扇贝进行攻毒感染实验。统计发现，具有-391 AA基因型的扇贝累计死亡率显著高于具有-391 AG基因型的扇贝（P=0.001），进一步证实了CfLysG基因-391 AG基因型与栉孔扇贝对鳗弧菌的抗性显著相关。CfLysG基因的外显子共有3处SNP，其中仅第三外显子上的+3473 A/C为非同义突变。统计分析表明，+3473位点不同基因型在敏感群体中的分布频率符合Hardy-Weinberg平衡（P>0.05），而在抗病群体中则偏离Hardy-Weinberg平衡（P<0.01）。+3473 AA基因型在抗病群体中的频率显著高于在敏感群体中的频率（P=0.022），表明+3473 AA基因型与栉孔扇贝对鳗弧菌的抗性显著相关。CfLysG基因第1内含子存在+96 I/D和+487 I/D两处大片段的I/D多态性。统计发现，这两个位点的基因型在敏感群体和抗病群体中的分布频率均符合Hardy-Weinberg 平衡（P>0.05）。其中+96 DD基因型和+487 ID基因型在抗病群体中的频率均略高于在敏感群体中的频率，但两者之间无显著性差异（P>0.05）。表明这两个位点的多态性与栉孔扇贝对鳗弧菌的抗性无显著相关性。对CfLysG基因各多态性位点的统计分析表明，各位点之间存在不同程度的连锁不平衡，提示有单体型的存在。对19种频率>1%的单体型在敏感群体及抗病群体中的频率进行分析，发现-753 I/-391 G/-284 I/+96 I/+487 D/+3473 A单体型在抗病群体中的频率显著高于敏感群体（P=0.044），表明该单体型与栉孔扇贝对鳗弧菌的抗性显著相关。 在栉孔扇贝CfC1qDC基因cDNA序列上共发现14处SNP。对+423 T/C多态性与栉孔扇贝对鳗弧菌抗性的关系进行了分析。统计发现，+423位点各基因型在敏感群体和抗病群体中的分布均符合Hardy-Weinberg平衡（P>0.05）。+423 TT基因型在抗病群体中的频率显著高于在敏感群体中的频率（P=0.005），表明+423 TT基因型与栉孔扇贝对鳗弧菌的抗性显著相关。 在栉孔扇贝CfLITAF基因cDNA序列中共发现3处SNP及1处I/D多态性。对+145 I/D多态性进行研究，发现所有敏感个体及抗病个体中均同时存在+145 位点所有等位基因，表明+145位点多态性与栉孔扇贝对鳗弧菌的抗性不相关。 以上研究表明，栉孔扇贝CfLysG基因-391 AG基因型、+3473 AA基因型、-753 I/-391 G/-284 I/+96 I/+487 D/+3473 A单体型以及CfC1qDC基因+423 TT基因型与栉孔扇贝对鳗弧菌的抗性显著相关，提示它们可作为与栉孔扇贝抗病相关的候选分子标记应用于贝类抗病育种中，为贝类的标记辅助育种提供参考。此外，抗病相关分子标记的发现还有利于加深对扇贝发病机理的理解，并有助于发掘预防及治疗贝类疾病的新方法。

Isolation and identification of bacteria associated with the surfaces of several algal species

We conducted this study to assess the diversity of bacteria associated with the surfaces of algae based on 16S rDNA sequence analyses. Twelve strains of bacteria were obtained from the surfaces of the following four species of algae: Gracilaria textorii, Ulva pertusa, Laminaria japonica, and Polysiphonia urceolata. The isolated strains of bacteria can be divided into two groups: Halomonas and Vibrio, in physiology, biochemical characteristics and 16S rDNA sequence analyses. The phylogenetic tree constructed based on 16S rDNA sequences of the isolates shows four obvious clusters, Halomonas venusta, Vibrio tasmaniensis, Vibrio lentus, and Vibrio splendidus. Isolates from the surface of P. urceolata are more abundant and diverse, of which strains P9 and P28 have a 16S rDNA sequence very similar (97.5%-99.8%) to that of V. splendidus. On the contrary, the isolates from the surfaces of G textorii, U. pertusa and L. japonica are quite simple and distribute on different branches of the phylogenetic tree. In overall, the results of this study indicate that the genetic relationships among the isolates are quite close and display a certain level of host species specificity, and alga-associated bacteria species are algal species specific.

Grazing impact of copepods on phytoplankton in the Bohai Sea

During spring (April/May 1999) and autumn (September/October 1998) cruises in the Bohai Sea, China, copepods were the dominant components of mesozooplankton, the most abundant species being Calanus sinicus, Centropages mcmurrichi, Paracalanus parvus, Acartia bifilosa and Oithona similis. Pigment ingestion rates by three size classes of copepods (200-500, 500-1000 and > 1000 mum) were measured. In the south of the investigation area, gut pigment content (GPC), individual pigment-specific ingestion rates and grazing impacts on phytoplankton were lower in spring than in autumn. In the central area, GPC and individual pigment-specific ingestion rates were higher in spring than in autumn. The grazing impact on phytoplankton by the copepod assemblages was lower in spring than in autumn, however, because of the relatively smaller biomass in spring. In the western area where the Bohai Sea joins the Yellow Sea, GPC, individual pigment-specific ingestion rates and grazing impacts on phytoplankton were higher in spring than in autumn. Among the three size groups, the small-sized animals (200-500 mum) contributed more than 50% (range 38-98%) of the total copepod grazing during both cruises. The grazing impact on phytoplankton by copepods was equivalent to 11.9% (range 3.0-37.1%) of the chlorophyll-a standing stock and 53.3% (range 21.4-91.4%) of the primary production during the spring cruise. Grazing impact was equivalent to 6.3% (range 2.0-11.6%) of the chlorophyll-a standing stock and >100% (range 25.7-141.6%) of the primary production during the autumn cruise. The copepod community apparently consumed only a modest proportion of the standing stock of phytoplankton during spring and autumn blooms. They did, however, sometimes graze a significant proportion of daily primary production and hence were presumably able to limit the rate of further accumulation of phytoplankton, or even to prevent it. (C) 2003 Elsevier Ltd. All rights reserved.

Immune condition of Chlamys farreri in response to acute temperature challenge

The effects of acute temperature challenge on some immune parameters of haemocyte in Zhikong scallop, Chlamys farreri, recognised as a temperature sensitive bivalve species, were evaluated over a short period of time. Scallops were suddenly transferred from 17 degrees C to 11 degrees C, 23 degrees C and 28 degrees C for a period of 72 h. Total haemocyte count (THC), percentage of phagocytic haemocytes, reactive oxygen species (ROS) production, acid phosphatase (ACP) and superoxide dismutase (SOD) activities (in both haemocyte lysate and cell-free haemolymph) were chosen as biomarkers of temperature stress. Results demonstrated that the percentage of phagocytic haemocytes and ACP activity in cell-free haemolymph of scallops challenged at 28 degrees C for 72 h significantly decreased. By contrast, reactive oxygen species production by haemocytes increased when compared to the initial values. It is concluded that haemocyte activities of C. farreri appear to be compromised when scallops were transferred from 17 degrees C to 28 degrees C. Meanwhile, no obvious negative effect of acute temperature stress was detected on haemocyte activities of C. farreri challenged at 11 degrees C, which highlighted the high tolerance of scallops to acute decrease of seawater temperatures. (C) 2007 Elsevier B.V. All rights reserved.

Identification of Crassostrea ariakensis and related oysters by multiplex species-specific PCR

Genetic markers are needed for rapid and reliable identification of oysters. In this study, we developed multiplex genus- and species-specific PCR markers for the identification of oysters from China. We used the mitochondrial cytochrome oxidase I (COI) and nuclear 28S ribosomal RNA genes for marker development. DNA sequences from different species were obtained from GenBank or by direct sequencing. Sequences were aligned, and genus- and species-specific nucleotides were identified. Primers were designed for genus/species-specific amplification to generate fragments of different sizes. A multiplex set of genus- and species-specific primers from the 28S gene was able to separate C. ariakensis and C. hongkongensis from other species and assign oysters to four genera. A set of species-specific COI primers provided positive identification of all five Crassostrea species from China, C. ariakensis, C. hongkongensis, C. angulata, C. gigas, and C. sikamea in a single PCR. The multiplex PCR assays do not require fluorescence-labeling or post-PCR enzyme digestion, providing a simple, fast and reliable method for the identification of oysters from China.

高寒草甸不同草地群落物种多样性与生产力关系研究

生态系统的结构和功能、生物多样性与生产力的关系问题是近年来群落生态学中研究的中心问题,其中,生态系统生产力水平是其功能的重要表现形式,用4种不同草地类型探讨自然群落的物种多样性与生产力关系.结果表明,矮嵩草草甸、小嵩草草甸和金露梅灌丛群落中物种多样性与生产力的关系呈线性增加关系,藏嵩草沼泽化草甸群落中线性增加关系不显著,这表明群落生产力除受物种多样性的影响外,也受物种本身特征和环境资源的影响.不同的环境资源和环境异质性是形成群落结构特征、物种多样性分布格局差异的主要原因之一.

青海省大黑沟种植大黄中矿物质元素研究

青海省地道中药材"西宁大黄"野生资源濒临枯竭,人工种植规模不断扩大.定期采集青海省大黑沟2年生种植大黄,分析测试其矿物质元素,为西宁大黄资源的可持续开发和利用提供科学依据.研究表明:大黑沟种植大黄的根、茎、叶等不同的组织器官,以及在不同生长季节对矿物质元素营养的需求具有动态性变化的特征.

贵阳市表层土壤中汞的环境地球化学基线及污染研究

以土壤环境地球化学研究为主线,在贵州省贵阳市8 046 km2研究区域中,将土壤重金属元素汞的空间分布规律与环境地球化学机理研究相结合,建立区域土壤环境地球化学基线,选用合适的判别指标判识自然作用过程与人类活动过程对土壤环境的影响,对487个样品的汞含量进行了统计分析.结果表明,贵阳市表层土壤中重金属元素汞的基线值为0.045 mg·kg^-1,样品中汞元素含量大于0.530 mg·kg^-1的样品可能遭受人为污染的影响.地质累积指数分析结果显示,贵阳市12%的表层土壤未受汞的污染,37%的表层土壤在无污染到中度污染之间,36%的中度污染,11%的表层土壤介于中度污染到强污染之间,2%的强污染,1%的介于强污染到极强污染之间.污染程度指数分析显示,贵阳市56.1%的表层土壤未受到汞的污染,汞的污染程度最大为45.87,总污染程度大于0（即受到污染）.

贵阳市表层土壤中镉的环境地球化学基线研究

以土壤环境地球化学研究为主线,以贵州省贵阳市8 046 km^2为研究区域,将土壤重金属污染元素镉的空间分布规律与环境地球化学机理研究相结合,建立区域土壤环境地球化学基线,选用合适的判别指标判识自然作用过程与人类活动过程对土壤环境的影响.通过对487个样品的镉含量的统计分析,结果表明,贵阳市表层土壤中镉的基线值为0.068 mg/kg,镉元素含量大于1.010 mg/kg的样品可能遭受人为污染的影响.地质累积指数分析结果显示,贵阳市40%的表层土壤未受镉污染,19%的表层土壤在无污染与中度污染之间,14%受中度污染,19%的表层土壤介于中度污染到强污染之间,7%受强污染,1%的表层土壤介于强污染到极强污染之间.污染程度指数分析则显示,贵阳市57.9%的表层土壤未受到镉污染,镉的污染程度最大为12.1,96%的表层土壤污染程度小于4,总污染程度大于0,即总体受到污染.[

云南会泽铅锌矿田成矿物质来源：Pb、S、C、H、O、Sr同位素制约

云南会泽铅锌矿田是我国著名的超大型特富铅锌矿田之一，由相距3公里的矿山厂和麒麟厂两个独立的矿床组成，Zn＋Pb金属量超过五百万吨，矿石品位在25%至35%之间。为确定矿床成矿流体和成矿金属来源，本文系统研究了矿床的Ph、S、C、O、H和Sr同位素组成特征。矿石硫化物的铅N位素组成均一，^206Pb／^204Pb，^207Pb／^204Pb和。^206Pb／^204Pb的变化范围分别为18．251～18．530，15．663～15．855和38.487～39．433，与围岩碳酸盐岩中浸染状黄铁矿一致，与碳酸盐地层相近，在^208Pb／^204Pb-^206Pb／^204Pb图上显示明显的线性关系，表明铅N位素组成相近的碳酸盐围岩地层提供了成矿物质。矿石硫化物的δ^14S变化范围为10．9‰～17．4‰，多数集中于13‰～17‰，表明还原硫主要来自地层中海相硫酸盐的还原，还原方式为热化学还原，下伏页岩、碎屑岩和泥质岩中的有机质在硫酸盐还原过程中发挥了重要作用。三种不同产状的脉石矿物方解石的碳氧同位素组成均一且没有明显差别，δ^13C变化范围为-2．1‰～-3．5‰, δ^18O为16.8‰～18．6‰。脉石矿物方解石中流体包裹体水的δDFI为-50‰～-60‰，取温度为200℃计算包裹体水的δ^18OH2O值为7．0‰～8.8‰。碳、氧和氢、氧同位素研究结果表明，成矿流体为来自下部上升的变质流体，由于下伏页岩、碎屑岩和泥质岩中有机质的参与，成矿流体具有低的δ^13C和δ^13DFI值，在上升过程中与围岩发生了同位素交换。矿石中黄铁矿、闪锌矿和方解石的初始锶同位素组成（^87Sr／^86Sr）．值的变化范围为0．714～0．717，赋矿围岩中未蚀变白云岩的初始锶同位素组成（^87Sr／^86Sr）．值为0.7083～0．7093，明显低于蚀变白云岩（0．7106），表明成矿流体具有高的（^87Sr／^86Sr），比值。相对围岩碳酸盐岩而言，下伏地层中的页岩、碎屑岩和泥质岩往往具有高得多的^87Sr／^86Sr，因此，流经或者起源于这些地层的流体具有高的锶同位素比值.

分散元素的超常富集与共生

本文以地质事实为基础，初步探讨了分散元素的超常富集和共生问题。在一定地质条件下，分散元素可以发生超常富集，乃至形成分散元素的独立矿床。低温条件有利于分散元素的超常富集和形成独立矿物；热水沉积作用在某些分散元素的成矿过程中具有重要意义。分散元素共生，是一种常见的现象。相似的地球化学 性质和地球化学行为，是导致分散元素共生的基本原因。男外，区域地质背景也是一个重要因素。

Enhancement of molar heat capacity of nanostructured Al2O3

The nanosized alumina prepared by the hydrolysis method with an average particle size of 20 nm was characterized by X-ray diffraction. The heat capacity measurements of the prepared sample were carried out using an adiabatic calorimeter in the temperature range from 78 to 370 K. Enhancement of heat capacity was observed in the nanostructured materials as the heat capacity data were compared with those of the corresponding coarse-grained materials. The enhanced heat capacity was discussed on the basis of experiments. Differential scanning calorimetry and thermogravimetry were used to determine the thermal stability of the nanostructured alumina.

Characterization of iron atoms in the framework of MFI-type zeolites by UV resonance Raman spectroscopy

Silicalite-I, ZSM-5, and Fe-ZSM-5 zeolites prepared from two different silicon sources are characterized by UV resonance Raman (UVRR) spectroscopy, X-ray diffraction (XRD), electron spin resonance (ESR), and UV/visible diffuse reflectance spectroscopy (UV/Vis DRS). A new technique for investigating zeolitic structure, UV resonance Raman spectroscopy selectively enhances the Raman bands associated with framework iron atoms incorporated into MFI-type zeolites, and it is very sensitive in identifying the iron atoms in the framework of zeolites, while other techniques such as XRD, ESR, and UV/Vis DRS have failed in uncovering trace amounts of iron atoms in the framework of zeolites. (C) 2000 Academic Press.

Imaging of three-dimensional objects in emission electron microscopy

Under investigation by emission electron microscopy, the shape and size of three-dimensional objects are distorted because of the appearance of a characteristic potential relief and a possible contact potential difference between the particles and the substrate. An estimation of these effects for spherical particles is made. It is shown that the apparent size of particles observed in an emission electron microscope (EEM) could be increased as well as decreased depending on the relation between the work functions of the particle and the substrate. The corresponding formulae are given and several possibilities are shown which permit us to determine from the EEM image the real size of particles and their work function relative to the substrate.