Heat-shock protein 70 expression in shrimp Fenneropenaeus chinensis during thermal and immune-challenged stress

Using western immunoblotting, we obtained heat-shock protein 70 (HSP70) induction data and distribution in different tissues from shrimp Fenneropenaeus chinensis during thermal and immune-challenged stresses. This is probably the first report of the effects of various stressors on the expression of HSP70 in shrimp.HSP70 was prominently induced in hepatopancreas and gills, but not in muscle, eyestalk and hemolymph, when the shrimp were exposed to heat shock and Vibrio anguillavium-challenged stresses. Cold shock and WSSV treatment had no significant effects on the levels of HSP70 expression in all tissues examined. HSP70 induction was greatest after 2 h exposure to heat shock stress, which was elevated after acute heat shock exposure of 10℃ above ambient temperature.     

富含半胱氨酸肠蛋白的研究进展

富含半胱氨酸肠蛋白是一种含双锌指结构的小分子蛋白，其在锌离子转运、个体发育、免疫防御以及肿瘤发生等多种生理过程中发挥重要作用。作者主要对富含半胱氨酸肠蛋白的结构特点及其在发育和免疫方面的研究进展进行系统概述，并指出其相关作用机制尚待深入研究。     

大黄鱼(Pseudosciaena crocea)转铁蛋白(Tf)基因克隆及序列分析

提要应用RNeasy animal mini试剂盒抽提大黄鱼total RNA,逆转录合成模板cDNA;同时综合分析从GenBank数据库查询到已报道的转铁蛋白(Tf)基因序列,先设计并合成扩增引物扩增出大黄鱼转铁蛋白部分基因序列,再应用RACE技术,克隆出大黄鱼转铁蛋白全长cDNA基因,全长2461个核苷酸,编码661个氨基酸。应用WCG软件包,采用DNADIST和NEIGHBOR分析方法,构建了转铁蛋白系统进化树。树图显示与传统的分类地位大致相符,可以看出转铁蛋白在海水鱼和淡水鱼的进化上的差异,同时也显示大黄鱼Tf与真鲷的同源性最高。     

96孔酶标板法测定中国对虾血淋巴上清液抗菌活力和酚氧化酶活性的初步研究

用96孔酶标板法测量单尾中国对虾血淋巴上清液抗菌（Uα）应力和过氧化酶活性。实验表明，该法具有以下优点：1）可测定大批单尾对虾血淋巴上清液的抗菌活力，酚氧化酶活性；2）所需对虾血淋巴上清液微量化；3）用酶标仪读取酶的活性，减少了不同试验者人为读数时的误差；4）重复性高。     

口服免疫药物后中国对虾某些血淋巴因子的测定及方法研究

于１９９１年４－９月，以海捕中国对虾亲虾及养殖中国对虾为材料，运用单向免疫扩散（ＳＲＩＤ）方法测定对虾血淋巴免疫因子，并采用经改进后的Ｈｏｒｏｗｉｔｚ等人的方法测定对虾体内的酚氧化酶（ＰＯ）活性，目的在于运用该两种方法对口服不同免疫药物的养殖对虾的血淋巴免疫因子及ＰＯ活力进行测定。对亲虾和养殖对虾口服不同免疫药物前后的测定结果表明，几种情况下的对虾血淋巴中均存在类似ＩｇＭ的因子，血细胞中存在酚氧化     

6-磷酸山梨醇脱氢酶(gutD)基因克隆、测序和表达

本研究根据6-磷酸山梨醇脱氢酶基因(gutD)两端序列设计引物，以假单胞杆菌(Pseudonmonas XM)的总DNA为模板通过PCR的方法克隆gutD基因并进行序列分析，将克隆得到的gutD基因与原核表达载体pBV220连接并转化大肠杆菌(Escherichia coli)JM101后检测其蛋白表达及菌株生长耐盐性。     

斑鳢(Channa maculata)微囊藻毒素去毒相关基因sGST的克隆与序列分析

采用RT-PCR技术和RACE技术成功克隆淡水鱼类斑鳢sGST基因cDNA全序列,推测得到氨基酸序列,初步分析其结构功能域及系统进化关系。结果表明,斑鳢sGST基因cDNA序列全长为898bp,编码225个氨基酸。斑鳢sGST与真鲷、金头鲷、鲽、黑头鲦、川鲽、大口黑鲈等最新定名为ρ型的sGST氨基酸同源性较高,ρ型sGST为水生生物所特有并共同占据进化树上独立的分枝;与大鼠、小鼠、人等哺乳动物sGST现有所有类型同源性均很低,并且在进化树上距离也较远,表明本研究成功克隆的sGST基因应属于ρ型,可能在鱼类等水生生物对水栖环境的适应上有重要作用。     

Rapid detection of virulent protease secreted by Vibrio anguillarum by dot enzyme-linked immunosorbent assay

Dot enzyme-linked immunosorbent assay （dot-ELISA）, indirect ELISA and Westem blot were performed to detect the virulent protease secreted by Vibrio anguillarum which was isolated from the diseased left-eyed flounder, Paralichthys olivaceous. Sensitivity results showed that dot-ELISA is a more sensitive, rapid and simple technique for the protease detection. The minimal detectable amount of protease is about 7 pg in the dot-ELISA test, while 7.8 ng in the indirect ELISA and 6.25 ng in the Westem blot respectively. Protease could be detected 2 h after incubation of V. anguillarum in the 2216E liquid medium but enzyme activity was very low at that period. From 6 to 12 h, the amount and enzyme activity of protease increased markedly and reached maximum at stationary phase. Analysis of serum samples periodically collected from the infected flounders showed that after 2 h of infection by V. anguillarum, the pathogenic bacteria could be detected in the blood of the infected flounders but no protease was found. It was 5-6 h after infection that the protease was detected in blood and then the amount increased as infection advanced. Quantitative detection of protease either incubation in the medium or from the blood of infected flounders could be accomplished in virtue of positive controls of quantificational protease standards （＂marker＂） so that the alterations ofprotease secretion both in vitro and in vivo could be understood generally. In addition, the indirect ELISA and dot-ELISA were also performed to detect V. anguillarum cells. Results indicated that the sensitivity of indirect ELISA to bacteria cells is higher than that of the dot-ELISA, and that the minimal detectable amount is approximately 10^4 cell/mL in the indirect ELISA, while 10^5 cell/mL in the dot-ELISA.     

海带几个不同种的雌性克隆对高温的适应力

每几个不同种的雌性克隆对高温(22℃、24℃、26℃)的适应力有所差异。大西洋沿岸的Laminariahyperhoren和L.sacchrina雌性克隆较太平洋沿岸的L.japonica、L.angustata和L.ochotensis雌性克隆明显地不耐高温,遗传性的不同是造成这种差异的主要原因。这种遗传性差异是海带长期自然选择的结果。同一物种L.japonica和L.japonicaCⅡ两种雌性克隆在24℃条件下,在第24天其死亡率分别是40.2％和25.2％,X~2=12.25,P<0.001,差异是高度显著的,说明这种不同是遗传性不同所致,而且属于物种内部的差异。     

中国对虾血细胞及其免疫研究

于１９９１年３月－１９９２年４月青岛近海及一养虾场捕集中国对虾亲虾和越冬亲虾，以扫描电镜和透射电镜观察中国虾的血细胞形态及其亚显微结构；研究定量测定细胞吞噬活性的方法。结果表明，中国对虾血细胞分３种：无颗粒细胞，呈球形或梭形，胞质不含颗粒粒；小颗粒细胞及颗粒细胞，卵球形或椭球球形，胞质含有电子致密颗粒，颗粒可分均匀、非均匀的；小颗粒细胞质富含线粒体，可能与它对异物敏感易发生胞吐作用有关，该细胞是识