Expression of putative zinc-finger protein lcn61 gene in lymphocystis disease virus China （LCDV-cn） genome

An open reading frame (lcn61) of lymphocystis disease virus China (LCDV-cn), probably responsible for encoding putative zinc-finger proteins was amplified and inserted into pET24a (+) vector. Then it expressed in E. coli BL21 (DE3), and His-tag fusion protein of high yield was obtained. It was found that the fusion protein existed in E. coli mainly as inclusion bodies. The bioinformatics analysis indicates that LCN61 is C2H2 type zinc-finger protein containing four C2H2 zinc-finger motifs. This work provides a theory for functional research of lcn61 gene. Supported by High Technology Research and Development Program of China (863 Program, No. 2006AA100309)     

Adsorption of Heavy Metal Ions, Dyes and Proteins by Chitosan Composites and Derivatives- A Review

Chitosan composites and derivatives have gained wide attentions as effective biosorbents due to their low costs and high contents of amino and hydroxyl functional groups. They have showed significant potentials of removing metal ions, dyes and pro- teins from various media. Chemical modifications that lead to the formation of the chitosan derivatives and chitosan composites have been extensively studied and widely reported in literatures. The aims of this review were to summarize the important information of the bioactivities of chitosan, highlight the various preparation methods of chitosan-based active biosorbents, and outline its potential applications in the adsorption of heavy metal ions, dyes and proteins from wastewater and aqueous solutions.     

6种海洋致病性弧菌36kDa外膜蛋白特性分析

研究本着筛选和制备弧菌共同外膜蛋白亚单位疫苗,用十二烷基肌氨酸钠抽提并结合超速离心的方法提取了鳗弧菌(Vibrio anguillarum)、鱼肠道弧菌(V.ichthyoenteri)、溶藻弧菌(V.alginolyticus)、副溶血弧菌(V.parahaemolyticus)、哈维氏弧菌(V.harveyi)和创伤弧菌(V.vulnificus)6种海洋致病性弧菌的主要外膜蛋白, SDS-PAGE图谱比较发现这6种弧菌都存在36 kDa的外膜蛋白条带.通过电泳洗脱分离纯化这6种弧菌的36 kDa外膜蛋白,并制备了鳗弧菌菌株SJ060621的36 kDa外膜蛋白多抗.ELISA和Western-blot印迹结果显示,仅鳗弧菌SJ060621、S010610-1和哈维氏弧菌的36 kDa外膜蛋白存在免疫交叉反应;双向电泳分析了鳗弧菌SJ060621、S010610-1、哈维氏弧菌和溶藻弧菌的36 kDa外膜蛋白,2-DE图谱显示出鳗弧菌SJ060621和S010610-1均由3种等电点相近的蛋白组成,存在很高的同源性,2株鳗弧菌与溶藻弧菌和哈维氏弧菌在蛋白质组成数量和等电点上都存在较大的差异,结果说明通过主要外膜蛋白的分子量和免疫特性研究,可为筛选不同弧菌共同外膜蛋白的亚单位疫苗提供有利参考.     

蛋白质营养对高密度养殖凡纳滨对虾生长与免疫力的影响

在高密度养殖条件下,进行单因素随机设计动物试验.用5种饲料(蛋白质水平31%、35%、39%、43%、47%,以A～E组表示)分别投喂平均体质量6.2 g+0.2 g、平均养殖密度3.1 kg/m3的凡纳滨对虾(Litopenaeus vannamei),探寻蛋白质营养对虾生长、免疫、水质、抗胁迫的影响特征.结果表明:(1)中高蛋白质水平具有显著促进对虾生长的效果,随着蛋白水平的提高,特定生长率先增加后降低,饲料系数正好相反,D组两指标最佳,分另q为110.98%和2.54;C、D、E组差异不显著.(2)中高蛋白质水平有利于提高对虾多数免疫指标的活力,血淋巴中血细胞浓度、T-AOC活力、POD活力、总蛋白含量、白蛋白、血蓝蛋白含量,随着蛋白质水平提高先增加后降低,前5指标含量均以D组最高,比A组显著提高16.8%～33.9%;而血蓝蛋白含量C组最高,比A组提高15.0%.(3)高蛋白质水平有利于提高对虾SOD活力,也显著增强抗低盐胁迫的能力,但同时极显著加大了水环境中氨氮和亚硝氮的污染.(4)在我国北方集约化高密度养殖条件下,凡纳滨对虾中后期生长阶段适宜的饲料蛋白质营养水平为39%～43%.     

谷氨酰胺二肽对日本对虾血清生化指标、肝胰腺细胞凋亡及肠粘膜形态的影响

采用饲养对比试验方法,研究了谷氨酰胺二肽（Gin二肽）对日本对虾血清生化指标、肝胰腺细胞凋亡和肠绒毛的影响及对虾体健康的作用。结果表明,饲喂Gin二肽各组的PO、LSZ、ACP含量均显著（P〈0．05）高于对照组,添加0.5％和1．0％Gin二肽组的SOD和ALP含量也显著（P〈0．05）高于对照组：添加1．0％Gin二肽的TP含量显著（P〈0．05）高于其它各组。血清TP、PA、UN、CHO、PO、LSZ、SOD、ALP、ACP和LDH等各项指标均随Gin二肽添加量的提高而呈现增加趋势。试验建立了流式细胞术检测对虾肝胰腺细胞凋亡率的方法;肝胰腺细胞凋亡率随着日粮Gin二肽添加量的增加而显著降低（P〈0．05）。肠道切片显微观测结果表明,添加1．0％的Gin二肽能显著增加日本对虾肠绒毛高度（P〈0．05）。饲料中添加Gin二肽可显著提高日本对虾血清中的溶菌酶、抗氧化酶及磷酸酶活性,降低肝胰腺细胞凋亡率,增加肠绒毛高度,改善虾体健康状况。     

Cloning and sequence analysis of a full-length cDNA of <Emphasis Type="Italic">SmPP1cb</Emphasis> encoding turbot protein phosphatase 1 beta catalytic subunit

Reversible protein phosphorylation, catalyzed by protein kinases and phosphatases, is an important and versatile mechanism by which eukaryotic cells regulate almost all the signaling processes. Protein phosphatase 1 （PP1） is the first and well-characterized member of the protein serine/threonine phosphatase family. In the present study, a full-length cDNA encoding the beta isoform of the catalytic subunit of protein phosphatase l（PPlcb）, was for the first time isolated and sequenced from the skin tissue of flatfish turbot Scophthalmus maximus, designated SmPPlcb, by the rapid amplification of cDNA ends （RACE） technique. The cDNA sequence of SmPPlcb we obtained contains a 984 bp open reading frame （ORF）, flanked by a complete 39 bp 5＇ untranslated region and 462 bp 3＇ untranslated region. The ORF encodes a putative 327 amino acid protein, and the N-terminal section of this protein is highly acidic, Met-Ala-Glu-Gly-Glu-Leu-Asp-Val-Asp, a common feature for PP1 catalytic subunit but absent in protein phosphatase 2B （PP2B）. And its calculated molecular mass is 37 193 Da and pI 5.8. Sequence analysis indicated that, SmPPlcb is extremely conserved in both amino acid and nucleotide acid levels compared with the PPlcb of other vertebrates and invertebrates, and its Kozak motif contained in the 5＇UTR around ATG start codon is GXXAXXGXXATGG, which is different from mammalian in two positions A6 and G3, indicating the possibility of different initiation of translation in turbot, and also the 3＇UTR of SmPPlcb is highly diverse in the sequence similarity and length compared with other animals, especially zebraf＇lsh. The cloning and sequencing of SmPPlcb gene lays a good foundation for the future work on the biological functions of PP1 in the flatfish turbot.     

黄斑篮子鱼去毒相关基因的克隆与肝脏组成型表达分析

从基因水平探讨海洋鱼类对海洋藻毒素的去毒分子机理。采用RT-PCR法成功克隆了黄斑篮子鱼Siganus oramin肝脏I时相代谢酶细胞色素P450 1A(CYP1A)、II时相代谢酶alpha型谷胱甘肽S-转移酶(GSTA)和rho型谷胱甘肽S-转移酶(GSTR)、热休克蛋白70 (HSP70)、alpha 1型钠钾ATP酶(ATP1A1)及β-肌动蛋白(beta-actin, ACT)基因cDNA核心序列,序列分别长879 bp、582 bp、588 bp、660 bp、749 bp和554 bp。序列同源性分析发现,属鲈形目的黄斑篮子鱼CYP1A、GSTA和GSTR与同属鲈形目的牙鲆Paralichthys olivaceus、欧洲鲽Pleuronectes platessa、真鲷Pagrus major、鲤形目的斑马鱼Brachydanio rerio 相应氨基酸序列同源性较高,CYP1A和GSTA与非洲爪蟾(两栖类)、鸡(鸟类)、小鼠、大鼠和人(哺乳类)相应氨基酸序列同源性低,这可能与鱼类I、II时相去毒酶基因承担水环境毒素去毒代谢的特殊功能有关;而HSP70、ATP1A和β-肌动蛋白在鱼类、两栖类、鸟类、哺乳类中均有较高的同源性,这可能与这些基因在机体中承担的最基本的生命功能相关。应用半定量RT-PCR的方法,以β-肌动蛋白作为外参照,在指数期增长范围内分别得到了CYP1A、GSTA、GSTR、HSP70和ATP1A1 mRNA与β-肌动蛋白mRNA (%)的比值,确定黄斑篮子鱼肝脏去毒相关基因的组成型表达水平。其中,黄斑篮子鱼肝脏CYP1A、GSTA和GSTR基因组成型表达相对较高,HSP70和ATP1A1基因组成型表达相对较低,这可能与不同基因在黄斑篮子鱼海洋藻毒素去毒分子机理中承担的作用相关,为海洋藻毒素在海洋鱼类中的积聚及代谢去毒分子机制的研究提供了相关数据。     

Economic assessment of production of flour from Prosopis alba and P. pallida pods for human food applications.

This paper presents a detailed cost accounting of the processes involved in the production of flours from the mesocarps of the fruits of Prosopis alba and P. pallida to be used in human foods. The flours from both species are similar and contain 7–8% protein, 0·8–2% fat, 26–32% total dietary fiber and 48–59% sucrose. The data were based on actual production rates in use at the Universidad de Piura pilot plant using simple machinery. Machinery hourly charges were based on equivalent rental costs in order to avoid interest charge calculations. As opposed to other analyses which only included processing costs delivered to the plant, our analysis ascribed value to the owner of the trees in order to provide incentives to plant and care for the trees. The sensitivity analyses showed that the most economically important factor was the conversion efficiency of pods into flour. When a 40% conversion efficiency, such as that which has been obtained in both Peru and Argentina was obtained, and by-product credits for high fiber containing fraction were assumed, the wholesale selling price with a 5% marketing cost and a 40% profit direct costs was estimated to be $0·66 kg⁻¹ for Peru (using a $0·40 h⁻¹ labor rate ) and $1·76 kg⁻¹ for Argentina (using a $1·5 h⁻¹ labor rate).     

Seasonal Changes in the Biochemical Composition of the Edible Sea Urchin Paracentrotus lividus Echinodermata: Echinoidea in a Lagoonal Environment

Abstract. Monthly soluble and insoluble protein, carbohydrate, lipid and ash levels and energetic content were measured in tissues of Paracentrotus lividus adults living in a lagoonal environment over a 1-year period to increase our understanding of resource allocation to the different tissues. The biochemical composition of P. lividus individuals in this environment is similar to that observed in other environments. The gonad and gut contained a high level of proteins soluble or insoluble, a lesser amount of lipids and, finally, a low level of carbohydrates. The main component of the test was ash, followed by protein levels. The lipid and carbohydrate levels in the test were very low. Seasonal variability in the biochemical composition of the different body parts was observed. In the gonad, an increase in the level of lipids was recorded concomitant to an increase in gonad weight. The lipids and carbohydrates, after an initial increase, decreased with the maturation of the gonad. The biochemical variations observed in the gut seem to be linked to both the reproductive cycle and the sea urchin's trophic activity. Variability in the test was low and irregular for all of the observed biochemical components. As P. lividus is an important member of the benthic community in the Urbinu lagoon, this echinoid represents an important store of materials and energy within this lagoonal environment.     

三角帆蚌(Hyriopsis cumingii)珍珠钙代谢的周年性变化研究

采用生态调查和生理生化方法,对植片后的1龄三角帆蚌(Hyriopsis cumingii)进行了为期一年的调查研究,拟探讨珍珠形成和钙代谢的相关性。定期测定珍珠囊和珍珠的重量、直径及水温,同时取其鳃、围心腔和外套膜组织进行酸、碱性磷酸酶活性检测及钙含量等测定。结果表明,1龄植片后的三角帆其蚌珍珠囊和珍珠在一周年之中出现三次快速生长期。外套膜中的酸、碱性磷酸酶均在165d时升至最高。鳃组织中钙含量显著高于围心腔和外套膜。经Pearson相关分析显示,珍珠囊和珍珠的重量、直径分别与三种组织中的碱性磷酸酶活性及外套膜中的钙含量呈显著性正相关(P<0.05);同时,外套膜中的钙含量与三种组织中的碱性磷酸酶呈显著性正相关(P<0.05);水温与外套膜、围心腔中的钙含量呈显著性负相关(P<0.05),与三种组织中的酸性磷酸酶呈显著性正相关(P<0.05)。提示:鳃是三角帆蚌钙吸收、贮存和钙调节的重要器官,形成珍珠的钙一部分来源于循环系统,另一部分可由外套膜直接从环境中吸收,碱性磷酸酶对珍珠生长起着重要的调节作用。