假雄牙鲆不同组织中溶菌酶比活性的研究   总被引：6，自引：1，他引：5  

王宏田  徐永立  张培军 《海洋科学》2000,24(10):7-8

溶菌酶（Ｌｙｓｏｚｙｍｅ）是鱼类非特异性免疫系统的重要组成部分。据Ｉｎｇｒａｍ１９８０年报道 ,溶菌酶能够破坏细菌的胞壁结构 ,增强由补体介导的溶菌作用 ,同时对真菌、寄生物以及病毒也具有破坏作用 ,因此 ,溶菌酶对于鱼类抵抗各种病原体的侵袭具有重要意义 ,了解溶菌酶在鱼体内的分布 ,有助于更好地了解鱼类抵抗各种疾病的机制。以往的实验对溶菌酶在鱼体不同组织中的分布研究得较少 ,对于牙鲆体内不同组织中溶菌酶活力的比较则未见报道。本实验以１ａ龄假雄牙鲆为实验材料 ,研究溶菌酶在不同组织中的分布 ,并初步比较其比活性的差…  相似文献   

鳗弧菌感染牙鲆的组织学和免疫组织化学观察   总被引：5，自引：0，他引：5  

夏永娟  黄威权  姜国良 《海洋科学》2000,24(10):41-44

利用免疫组织化学ＡＢＣ法及组织学ＨＥ染色技术对鳗弧菌人工感染后的牙鲆肝组织及胃肠道进行了组织病理学及细菌的定位研究。结果显示,感染后肝细胞肿胀,空泡化现象严重,肝窦扩张,肝组织内可见阳性反应的细菌呈堆状存在;胃黏膜褶变矮。胃壁变薄,黏膜层内可见阳性反应的细菌;肠绒毛细胞出现破损,肠腔内有大量细胞碎片,肠腔内也可见阳性反应的细菌。提示：鳗弧菌可直接侵入组织细胞破坏组织结构,进而使鱼体出现死亡。  相似文献   

凡纳滨对虾翻译控制肿瘤蛋白Fortilin的毕赤酵母表达及其对血细胞免疫反应的影响     

周怡  张文兵  麦康森 《中国海洋大学学报(自然科学版)》2012,42(1)

翻译控制肿瘤蛋白Fortilin是1种多功能蛋白,参与重要的细胞活动.并且,对虾fortilin通过抑制病毒复制干扰病毒感染.参照Genbank中fortilin基因序列设计引物,PCR扩增得到凡纳滨对虾fortilin目的基因片段,EcoR I/XbaI双酶切插入表达载体pGAPZαA,转化大肠杆菌,经博来霉素(Zeocin)抗性筛选及测序分析,获得分泌型重组酵母表达载体pGAPZαA-F.酶切线性化后,经电穿孔法转入毕赤酵母细胞X-33,经Zeocin抗性筛选,得到阳性转化子.表达产物的上清液经SDS- PAGE电泳和质谱分析,表明在酵母中成功表达fortilin.以体外原代培养的对虾血细胞检测重组蛋白的免疫活性,结果显示重组蛋白显著提高了血细胞酚氧化酶和超氧化物歧化酶活性.该结果为下一步研究重组fortilin 在对虾养殖中的应用奠定了基础.  相似文献   

壳聚糖对虹鳟(Oncorhynchus mykiss)幼鱼生长性能、体组成及非特异性免疫的影响   总被引：2，自引：0，他引：2  

蒋锦坤  王际英  张利民  柳旭东  冯德智  姜柯君  张德瑞 《海洋与湖沼》2012,43(4):729-734

用壳聚糖添加量为0.00%(D0)、0.25%(D1)、0.50%(D2)、1.00%(D3)和2.00%(D4)的实验饲料投喂虹鳟幼鱼50d,研究其对虹鳟幼鱼生长性能、体组成及非特异性免疫的影响。结果显示,各实验组增重率(WGR)、特定生长率(SGR)显著高于对照组(P<0.05),饲料系数(FCR)、脏体比(VSI)呈显著降低趋势(P<0.05),而肝体比(HSI)、脾体比(SSI)、肥满度(CF)及存活率(SR)无明显差异(P>0.05)。全鱼、肌肉及肝脏粗脂肪含量显著降低(P<0.05)。D3组和D4组碱性磷酸酶(ALP)活性显著低于其它各组(P<0.05);酸性磷酸酶(ACP)活性显著降低(P<0.05),溶菌酶(LZM)活性则显著升高(P<0.05);D4组总抗氧化能力(T-AOC)显著高于对照组及D1组(P<0.05);超氧化物歧化酶(SOD)活力差异不显著(P>0.05)。在本实验条件下,饲料中添加壳聚糖可显著提高虹鳟幼鱼生长性能、增强非特异性免疫能力,以增重率及非特异性免疫为综合评价指标,虹鳟饲料中壳聚糖的适宜添加量为0.50%。  相似文献   

中国明对虾(Fenneropenaeus chinensis)丝氨酸蛋白酶同源物基因(Fc-SPH)的重组表达及活性分析     

杨燚  刘逸尘  张亦陈  孙妍  耿绪云  孙金生 《海洋与湖沼》2012,43(4):852-856

以实验室前期克隆到的中国明对虾丝氨酸蛋白酶同源物基因(Fc-SPH)(GenBank登录号:DQ318859)为基础,利用原核表达系统对Fc-SPH基因成熟肽区域进行了重组表达和纯化复性分析,并对获得的重组目的蛋白开展了抑菌活性及微生物清除功能研究。结果表明:在体外成功获得了大量有活性的对虾Fc-SPH蛋白(rFc-SPH),活性研究显示rFc-SPH对大多数受试菌种都有明显的抑制效果,并可以加速中国明对虾清除体内外源微生物的速度。作者推断Fc-SPH即可以作为一线防御应答效应物直接参与虾类的先天免疫活动而发挥作用,也可以通过调理作用促进血细胞对病原微生物的吞噬和杀灭。  相似文献   

Molecular Cloning and Expression ofPoIR2, a Novel Gene Involved in Immune Response in Japanese Flounder （Paralichthys olivaceus）     

Shuo Li  Chunmei Li  Xubo Wang  Yanan Wang  Zhipeng Liu  Teng Zhai  Quanqi Zhang 《中国海洋大学学报(英文版)》2010,9(1):59-64

A novel immune-related gene was expressed in Japanese flounder (Paralichthys olivaceus) injected with Vibrio anguillarum. The complete cDNA contained a 169 bp 5’UTR, a 336 bp open reading frame (ORF) encoding 111 amino acids and a 556bp 3’UTR. Six exons and five introns were identified in the PoIR2 gene. Blastp similarity comparison showed its encoding protein had 50% similarity to Danio rerio neuromedin S (NMS), but further alignment indicated they did not have NMS C-terminal conservational signature domain. So it was not defined as an NMS homologue. Protein structure analysis indicated it had a 26aa signal peptide and was a secretory pathway protein. RT-PCR demonstrated that the expression of PoIR2 was quickly induced and drastically increased in liver, kidney, spleen, gills, intestine, heart, and skeletal muscle after infected with V. anguillarum. These results indicated that the PoIR2 might play some important role in Japanese flounder immune response system. This gene was named PoIR2 (P.olivaceus immune-related gene 2, GenBank accession number: EU224372). The mature PoIR2 peptide was expressed in BL21(DE3) pLysS using pET-32a(+) vector and a great part of the recombinant mature peptide existed as soluble type.  相似文献   

Effects of Dietary β-Glucan and Glycyrrhizin on Non-Specific Immunity and Disease Resistance of the Sea Cucumber （Apostichopus japonicus Selenka）Challenged with Vibrio splendidus     

CHANG Jie  ZHANG Wenbing  MAI Kangsen  MA Hongming  XU Wei 《青岛海洋大学学报(英文版)》2010,(4):389-394

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核酸免疫及其在水产病害防治中的应用前景   总被引：1，自引：0，他引：1  

王雷 《广东海洋大学学报》1997,17(2):80-82

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Production of monoclonal antibody and development of an enzyme-linked immunosorbent assay for the determination of okadaic acid in shellfish from China     

Renyan LIU Bingjun CHEN Yubo LIANG Daoyan XU Bing LIANG 《中国地球化学学报》2006,25(B08):217-217

An indirect competitive enzyme-linked immunosorbent assay（idc-ELISA） for quantitation of okadaic acid （OA） ,a marine biotoxin associated red tide, was developed by preparation of the monoclonal antibody against OA using cell-fused method.OA was coupled to KLH and OVA using carbodiimide reaction. OA-KLH used as immunogen were injected into Balb/C mice. Titres of the antisera against OA were determinated using OA-OVA as coating ligand by ELISA method. The spleen cells of immunized mice were fused with Sp2/O cells. Clones secreting specific monoclonal antibody were screened by indirect ELISA. After cloning, three hybridoma cell clones stably producing anti-OA monoclonal antibody were obtained. The detection of OA concentrations by indirect competitive ELISA was established. Under excellent condition, the detection limit of OA was 0.781 ng/mL. The level of OA in shellfishes from China was 0.001. A good correlation （R=0.7963） was observed between idc-ELISA and HPLC-MS/MS.  相似文献   

Development of an enzyme-linked immunosorbent     

Renyan LIU Bingjun CHEN Yubo LIANG Daoyan XU Bing LIANG 《中国地球化学学报》2006,25(B08):217-218

The development of immunodetection method of 2, 2-Bis （4-chlorophenyl） acetic acid （DDA）, the decomposition product of organochlorine insecticide DDT, was performed in this study. DDA was conjugated with bovine serum albumin （BSA） and ovualbumin （OVA） for the use of immunogen to produce antibodies and coating ligands to measure the titration level of antibody and the displacement of free analytes. Evaluation of the antisera by antibody capture assay showed that the antibody titer of antisera raised against DDA-OVA was higher than 1 ： 80000. Using 5#PAb, an indirect competitive enzyme immunoassay was developed for measurement of DDA. The working range for quantitative measurement of DDA and the quantitative limit for DDA were estimated to be 2.5-2000 ng/mL and 2.5 ng/mL for seawater, 12.5 ng/g for shellfish, respectively.  相似文献