Potassium channel in peripheral blood lymphocytes of turbot (Scophthalmus maximus)

In order to provide pertinent evidence of ion channel with immune response in the fish,whole cell patch-clamp technique was employed for potassium ion channel study in turbot(Scophthalmus maximus) . Lymphocytes were isolated by Percoll density gradient centrifugation from peripheral blood samples,and electrophysiological characters of the channel were analyzed. In the recorded cells,activated voltage of the channels was-42.5 ± 3.7 mV and the average peak current was 313.12 ± 28.2 pA. The channel was identified as voltage dependent,the current was outward and it could be inhibited by 10 mmol/dm 3 TEA or 5 mmol/dm 3 4-AP,a specific potassium channel inhibitor,identifying the existence of potassium channel in peripheral lymphocytes of the turbot.     

α-葡萄糖苷酶抑制剂类药物的研究进展

糖生物学领域的不断发展逐步揭示出糖苷酶作为多种反应的催化剂在体内所起白守重要作用,目前已知的功能包括碳水化合物的消化、糖蛋白和糖脂的加工,尔后二者又涉及到免疫反应、肿瘤的转移、病毒和细菌的感染以及神经细胞的分化。因此糖苷酶不仅是生命体正常运转所必需的酶,同时又是许多疾病（如糖尿病、爱滋病及癌症）的相关酶。目前通过抑制酶活的方法进行疾病的治疗已经成为药物研究领域的一个热点。而对高专一性的糖苷酶抑制剂的研究不仅可以用于研究酶活性中心的结构、酶的作用机制、疾病的产生原因,同时也使它们有望成为许多疾病的治疗药物。     

太平洋牡蛎中Cg-IL-17和Cg-TGF-β的表达及其免疫作用的研究

目的:探讨Cg-IL-17和Cg-TGF-β在太平洋牡蛎中表达的组织定位、免疫作用及其可能的机制。方法:采用Real-time PCR、原位杂交和图像分析的方法进行研究。结果:实时定量PCR研究表明,Cg-IL-17和Cg-TGF-β在弧菌感染后的太平洋牡蛎的鳃、外套膜、心包液、唇须、消化道和闭壳肌中表达明显增强。原位杂交实验结果表明,Cg-IL-17和Cg-TGF-β在未染菌组牡蛎的消化道、消化腺、鳃、唇须、外套膜中都有表达;染菌组以上各组织中的Cg-IL-17和Cg-TGF-β表达量均有显著增高,而消化道、消化腺中的表达量明显高于其他组织,闭壳肌中也均有微弱表达;感染弧菌前后两种基因在性腺中均未见表达。相关性分析发现这两种基因表达量在唇须和外套膜中具有明显的相关性。结论:Cg-IL-17和Cg-TGF-β具有多种生物学效应,且都参与了太平洋牡蛎的固有免疫反应;消化道可能是牡蛎主要的免疫反应器官之一;Cg-IL-17和Cg-TGF-β在唇须和外套膜中可能存在调控关系或协同作用。     

魁蚶(Scapharca broughtonii)热休克蛋白90(HSP90)基因的克隆及转录表达分析

热休克蛋白90(HSP90)作为一种研究的常规免疫基因,在许多物种都有过报道。本研究从构建的魁蚶转录组文库中筛选得到的HSP90基因部分序列为基础,通过RACE技术获得其c DNA全长序列(命名为Sb HSP90),以期明确魁蚶HSP90基因的结构特征、组织分布及其对病原菌刺激的免疫变化规律。序列和结构分析表明,该c DNA全长2707bp,编码一个由728个氨基酸组成的多肽,该多肽含有HSP90家族共有的5个签名序列,C端高度保守的MEEVD短肽序列和ATPase结构域;预测蛋白的分子量(Mw)为83.72k Da,理论等电点(p I)为4.85;预测该蛋白无信号肽,具有4个糖基化位点。同源性及系统分析表明,Sb HSP90基因与软体动物的HSP90相似性达到83%以上,其中与长牡蛎(Crassostrea gigas)和海湾扇贝(Argopecten irradians)相似度最高达86%,与甲壳动物HSP90的相似度都在81%左右,与脊椎动物HSP90-α和HSP90-β的同源性都很接近。实时荧光定量PCR(q RT-PCR)结果表明:Sb HSP90 m RNA在魁蚶血细胞、斧足、鳃、外套膜、闭壳肌和肝胰腺和中均有表达,斧足中的表达量相对较高,而在肝胰腺中的表达量则相对较低;注射鳗弧菌后,相对于对照组,Sb HSP90基因在所检测的每个组织中m RNA水平上的表达量都显著上调(P0.01),而且具有显著的时间依赖性和瞬时表达趋势。     

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

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.     

流式细胞术检测脂多糖对罗氏沼虾血细胞活性和免疫功能的影响

1引言 革兰氏阴性菌是虾类的一大致病菌,深入了解虾类对革兰氏阴性菌的免疫防御机制以及革兰氏阴性菌对虾类的致病机理,对虾类健康养殖有着重要的科学和实践意义.