摘要:通过构建斜带石斑鱼垂体cDNA文库,克隆了其生长激素(GH)全长cDNA.斜带石斑鱼GH全长cDNA为955bp,编码的多肽为204 aa.应用PCR方法把编码GH成熟肽的cDNA片段克隆到表达载体pET-15b,在大肠杆菌BL21(DE3)表达N端含6个组氨酸的融合多肽.SDS-PAGE结果表明,0.4 mmol@L-1IPTG诱导表达的蛋白约为24 kDa,主要为不溶性的包含体.细菌裂解液沉淀溶于6 mol@L-1盐酸胍后,用Ni2 -NTA树脂进行亲和分离纯化,纯化产物在SDS-PAGE上表现为一条24 kDa的蛋白带.在黑鲷GH放射免疫分析系统中,纯化产物能与黑鲷GH竞争结合GH抗体,表明大肠杆菌表达的斜带石斑鱼GH融合多肽具有GH免疫活性.

摘要:采用Olympus 3?51相差系统显微镜和SQIAS-1000彩色精液质量图文分析系统检测黄鳝精子活力.结果表明,在NaCl溶液浓度为0～0.3%时,黄鳝精子激活比例随溶液浓度升高而极显著增加(P＜0.01);当NaCl浓度达到0.7%时,精子激活比例、直线运动速度和鞭毛摆动频率均显著(P＜0.05)或极显著(P＜0.01)降低.扫描电镜观察显示:黄鳝成熟卵卵壳膜上的精孔区呈漏斗状凹陷,其底部中央可见一精孔管外孔,口径约4.22?0.66 μm;黄鳝精子入卵速度缓慢,受精过程较长,从精子附着于卵球表面到精孔管完全堵塞,约30s～5min.

摘要:在优化RAPD检测条件的基础上,从100个随机引物中筛选出60个扩增较好的引物对异源四倍体鲫鲤及其原始亲本红鲫和湘江野鲤相互间扩增DNA片段的异同性进行比较研究.结果显示,60个引物共产生1 554条带,单一引物扩增的DNA片段数在6～20条之间,片段大小在200～3 500 bp之间.遗传相似率分析表明,异源四倍体鲫鲤与其原始母本红鲫的相似率为69.41%,与其原始父本野鲤的相似率为64.47%,说明异源四倍体鲫鲤接受原始母本红鲫的遗传物质要多一些;而红鲫和野鲤之间的相似率为42.18%,说明两者的基因组成有较大的相似性,从分子水平证实了红鲫和野鲤的远缘杂交具有较大的亲和性.在分析中,还发现异源四倍体鲫鲤有4条非亲本带,非亲本带谱率达0.72%,这些非亲本带可以作为异源四倍体鲫鲤特异性的分子标记.

摘要:通过体外重组,将绿色荧光蛋白基因编码序列克隆到鲤鱼肌动蛋白基因启动子下游,构建成能在鱼体内表达绿色荧光蛋白的重组分子.用限制性内切酶PvuI酶切线性化后,通过显微注射法导入金鱼受精卵内,在转化后48h,可观察到绿色荧光,经PCR初步筛选后,对显阳性个体进行Southern杂交检测,结果表明,转化个体表现出较强的杂交信号.这说明绿色荧光蛋白基因能在金鱼体内整合、表达.

摘要:α2巨球蛋白能结合并抑制蛋白酶的活性,但不抑制该蛋白酶对人工合成的小分子底物的酶活.利用这一特性,选择胰酶的一种有色小分子底物BAPNA,建立BAPNA比色检测法,对7种常见养殖鱼:草鱼、鲫、鲢、胡子鲇、鳙、加州鲈和黄鳝的血清胰酶抑制活性进行了比较,胰酶抑制活性由高到低依次为:草鱼＞鲫＞黄鳝＞鳙＞鲢＞胡子鲇＞加州鲈;α2巨球蛋白活性由高到低依次为:黄鳝＞草鱼＞胡子鲇＞加州鲈＞鲢＞鳙＞鲫;黄鳝和草鱼的α2巨球蛋白活性明显高于其他几种鱼的α2巨球蛋白活性.

摘要:根据22头日本三陆沿岸白胸拟鼠海豚的检测结果,研究了必需元素铁在13种组织和器官中的蓄积特征.肺、肝脏和脾脏中铁的浓度显著高于其它脏器,铁在皮肤和鲸脂中的浓度却非常低.研究发现,鲸类肝脏中铁的平均浓度与其最大潜水持续时间有着极其显著的正相关(y=285.17x 0.5267;r2=0.92;y:a肝脏中铁的平均浓度,单位为μg@g-1干重;x:最大潜水持续时间,单位为min).利用该关系式,首次推测白胸拟鼠海豚的最大潜水持续时间为11.85min.

摘要:以皱纹盘鲍的血细胞为材料,采用植物血球凝集素(PHA)体外浸泡法,运用正交实验设计,探讨PHA处理时间、PHA浓度、抽血时间对细胞分裂频度的影响.根据正交实验直观分析结果,得出用血细胞直接制备染色体标本进行三倍体皱纹盘鲍活体倍性检测的最优组合:PHA处理时间18h、PHA浓度0.04%,抽血时间22:00-24:00,3因素的主次顺序:PHA处理时间→PHA浓度→抽血时间.并可获得大量图象清晰、长度适中、分散良好的中期分裂相,是皱纹盘鲍活体倍性检测的最佳方法.同时,通过流式细胞仪测定皱纹盘鲍血细胞DNA相对含量进行三倍体皱纹盘鲍活体倍性检测,该方法制样简单,测试速度快.

摘要:通过测定美国奥利亚罗非鱼(Ao)、Ao(♀)?鳜(♂)的子代(AsI)、埃及奥利亚罗非鱼(Eo)、Eo(♀)?鳜(♂)的子代(Es1)、As1(♀)?Ao(♂)的所谓"回交"子代(As2)、Es1的兄妹交子代(Es2)的形态和框架数据,用卡方分析、聚类分析、判别分析,比较了它们之间的形态异同.可数性状卡方分析表明,这几组实验鱼可数性状无显著差异;形态和框架数据的聚类分析结果表明,除了As2与Ao之间无显著差异外,其它都有显著差异,其中As1和Es1与其它鱼的差异最大,Es2与其它鱼的差异其次;判别分析结果表明,判别效果极显著,判别准确率都在90%以上,判别准确率依次为Es1、As1、As2都为100%,Es2为96.2%,Ao为95%,Eo为90%.综合以上分析,根据亚种75%规则,表明雌性奥利亚罗非鱼与雄性鳜间远缘杂交后代与亲本奥利亚罗非鱼不同;为此通过本课题的远缘杂交也许能产生新的养殖品种.

摘要:氏沼虾是我国从斯里兰卡新引进的淡水虾类,外观略呈纺锤形,虾体共分为20节,除尾节外,虾体的每一节各具1对附肢,雌虾和雄虾在第五步足基部分别具有"Λ"和"M"状突起.头胸甲的胃区、心区、触角区表面较粗糙,两侧还有数条与虾体平行的浅棕色斑纹.虾苗时期多数个体呈淡褐色,性成熟后体色呈浅褐、淡青偏褐色或淡灰色.额角尖端微微上翘向前伸展,但不超出第二触角的触角片末端,上翘高度不超过额角上缘鸡冠状隆起的顶端,齿式为7～13/4～6.其生物学特性与罗氏沼虾和日本沼虾相比有许多相似之处,但在体色、额角、头胸甲和腹甲侧片等形态特征上差异十分明显.本文描述了马氏沼虾的外部形态及结构,比较了上述几种虾的主要特征,并就"雄虾特化"现象以第二步足与体长之比作为依据提出了质疑.

摘要:根据室内流水实验法,定量研究了黄、渤海生态系统7种鱼类能量收支组分,并建立了相应的生物能量学模式.7种海洋鱼类的生物能量学模式显著不同,并可分为3类:(1)较低代谢和较高生长,如黑??、矛尾( )虎鱼;(2)较高代谢和较低生长,如欧氏六线鱼和黑鲷;(3)代谢和生长均处于中等水平,如真鲷、红鳍东方??和鲐.其差异原因可能与这7种海洋鱼类的生态习性不同相关.与淡水肉食性鱼类比较,7种海洋鱼类的代谢能明显偏高,表明海洋鱼类属于高代谢消耗、低生长效率型鱼类.

摘要:以白鱼粉为蛋白源设计了6个不同蛋白质水平(35%,40%,45%,50%,55%,60%)的等能饲料,采用室内水泥池养殖,在水温为24～26℃条件下对暗纹东方鲀幼鱼进行了12周的生长实验以确定其最适蛋白质需要量.增重率和饲料效率随着蛋白质水平的增加而上升,50%试验组的鱼体增重显著高于35%、40%和45%试验组(P＜0.05),蛋白质含量高于50%以后增重不再有显著变化(P＞0.05).蛋白质效率总体上随着蛋白质水平的增加趋于降低,但在35%、40%和50%试验组之间无显著差异(P＞0.05).采用折线模型测算出暗纹东方鲀幼鱼获得最大增长和对饲料利用最佳时的蛋白质水平分别大约是49%和46%,据此估算出暗纹东方鲀幼鱼的最适蛋白质含量为46%～49%,这一结果同红鳍东方鲀的最适蛋白质需要量基本一致. 关 键 词：暗纹东方鲀 蛋白质最适需要量 相对增重率 饲料效率 蛋白质效率 折线模型

摘要:采用高生物价的酪蛋白、明胶为蛋白源的蛋白饲料(PD)和无蛋白饲料(FPD)饲养南美白对虾幼虾(0.2627～0.2715g),根据虾体必需氨基酸(EAA)生长及维持量代谢,研究南美白对虾必需氨基酸的需求量.结果表明,要满足幼虾正常生长,各种EAA需求量[g@(100g虾体重)-1@d-1]为苏氨酸(Thr)0.046,缬氨酸(Val)0.054,蛋氨酸(Met)0.029,异亮氨酸(Ile)0.069,亮氨酸(Leu)0.087,苯丙氨酸(Phe)0.051,赖氨酸(Lys)0.086,组氨酸(His)0.025,精氨酸(Arg)0.097.如饲料蛋白质水平为40%,蛋白质利用率50%,虾摄食率为20%,推算得出饲料中EAA需求量(g@kg-1):苏氨酸(Thr)11.5,缬氨酸(Val)13.5,蛋氨酸(Met)7.3,异亮氨酸(Ile)17.3,亮氨酸(Leu)21.8,苯丙氨酸(Phe)12.8,赖氨酸(Lys)21.5,组氨酸(His)6.3,精氨酸(Arg)24.3.

摘要:利用组织学和组织化学方法,在光镜水平下研究了患"红肉病"文蛤(Meretrix meretrix Linnaeus)的组织病理学变化特征.结果显示,患病文蛤的病理学变化主要表现为组织结构紊乱,上皮膨大、脱落,鳃、外套膜、消化盲囊等组织发现异常结构及寄生物,如嗜碱性的包涵体、嗜酸性颗粒及寄生性原生动物等.另外,组织化学研究结果显示,病蛤在糖含量、磷酸酶活性等方面也有明显变化,表现为消化盲囊、肠等部位吸收细胞内糖含量增加.消化盲囊、消化管各处酸性磷酸酶(ACP)活性减弱,碱性磷酸酶活性(AKP)增强.鳃组织ACP活性增强,AKP活性减弱等.

摘要:通过体外试验,探讨了培养条件的改变对蟹源拟态弧菌(HX-4)产毒量的影响.结果表明,培养基种类、氯化钠浓度、初始pH、溶解氧、培养温度和时间的不同均可影响细菌产毒量,HX-4菌株的最佳产毒条件为细菌接种于起始pH为8.0,含0.5%NaCl-BHI培养基,28℃振荡培养36h或静置培养48h.同时,通过动物实验测定了外毒素的致病性,结果显示,外毒素粗提液对小鼠及河蟹均有致病性.粗提液的溶血活性越高,对动物的致死率也越高.具有蛋白酶活性而无溶血素活性的粗提液,对动物只有致病性,而无致死性.

摘要:将草鱼出血病病毒(GCHV)的结构蛋白与Sepherose-4B偶联制备亲和柱,经亲和层析从团头鲂血清中分离到一种蛋白,能凝集兔红细胞,其血凝性依赖于β巯基乙醇的存在.红细胞凝集抑制实验证实该蛋白与D半乳糖有最高的亲和力,其次为甘露糖.梯度PAGE测得其分子量约为240 000,SDS梯度PAGE证实该蛋白由分子量为15 000的单个亚基组成,亚基之间没有共价连接.以上特性与半乳糖凝集素相符,故称其为团头鲂抗病毒半乳糖凝集素样蛋白.抗体阻断试验证实,此种凝集素样蛋白是团头鲂血清抗病毒及红细胞凝集活性的主要成分.N末段的氨基酸序列为Lys-Val-Asn-Leu-Asp-Glu-Lys-Cys-Pro-Phe,检索GenBank,未发现与这一段序列同源的蛋白质.

摘要:以肌动球蛋白的盐溶性、肌原纤维蛋白ATPase活性以及肌原纤维蛋白的巯基含量为指标,研究了鳙肌原纤维蛋白在-10℃,-20℃,-30℃和-40℃下冻藏时的变性情况.结果表明,在不同温度下冻藏时,鳙肌动球蛋白的盐溶性、肌原纤维蛋白的ATPase活性以及巯基含量随着冻藏时间的延长,均呈下降趋势.但是,鳙蛋白质的变性速度在不同冻藏温度下的差异是极其显著的(P＜0.01).冻藏温度越低,变性越缓慢.

摘要:To observe the surface microstructure features by using scanning electron microscope , and study the surface antigen on lymphocytes of Paralichthys oliveaceus by form rosette with sheep red bloods cells and the cross-reactivity by using monoclonal antibody against human CD .Under scanning microscope , there are two kinds of lymphocytes , type I lymphocytes is characterized with being full of bumpiness and holes , the surface of type II lymphocytes is rather smooth with tiny finger like tubercle .The rosette formation of lymphocytes with sheep red blood cells is obvious however the rosette forming percents of lymphocytes is rather low, lower than 2 %.The cross reactivity of lymphocytes with monoclonal antibody against human CD showed that there are 5 .25 % 0 .38 % of NK cells in peripherial lymphocytes of Paralichthys oliveaceus , and the CD25 percentage of lymphocytes is 5 .25 % 0 .38 %.No cross-reactivity with T-cell antigen ,B-cell antigen and other activity antigen is found on the lymphocytes surface of Paralichthys oliveaceus .

摘要:A pair of primers was designed based on the vp 28 gene and PCR was performed to amplify the gene f rom WSSV DNA. Insert ing the DNA fragment to the yeas-t E . coli shuf fle vector pPICZ and the recombinant plasmid ( pPICZVP28) that contains the target DNA fragment was obtained in the E. coli strain. The pPICZVP28 was introduced into Pichia pastoris strain X33 by electroporation. The transformant strain X33-1 was grown in BMGY media in fled f lask at 28 e . Induced by methanol for 72h, the samples of cell pellets and supernatant were collected by centrifugation and analyzed by SDS-PAGE and Western-blot, which confirmed that the strain X33-1 can express the WSSV. s envelope protein vp 28.

摘要:Hard clams Mercenaria mercenaria were introduced from America to China in 2000. In this paper, the contents of 18 common acids, protein, fat as well as vitamines and mineral ions in edible part of hard calm were analyzed. The results showed that the contents of 18 common acids and the essential amino acid were 52. 42% and 40. 88% respectively and the average score of essential amino acid was 96. 4 .. 11. 3 according to the FAO/ WHO suggested level ( 1973) , and in which the smelled amino acid takes about 48. 30% . The content of protein in edible part of hard clam was higher( 60. 69% ) and the content of fat was much lower( 5. 60% ) than that of some bivalves. The results also indicated that the edible part of hard clam was also rich in amino acids, vitamins ( A, B1, B2, B5, B6) , and mineral ions such as Ca, Zn, Fe, Mn, etc. Hard clam was suggested to be with higher nutritional value which was good for human health.

摘要:When the temperature runs up to 20 .. , the male Oreochromis nilotica w ill excavate holes on the sides or the bottom of the pool. Oreochromis nilotica will hide in the holes when scared by the fishing haul or other provocat ion. This would make the fishing operation very difficult , and the f ishing rate very low . With the ordinary seine net, the fishing rate of f irst haul is always under 30% , and the total fishing rate for the first three hauls is about 50%. On the basis of the study on the behavior of Oreochromis nilotica, a new fishing gear, a seine with a curtain net , is designed. On the newly designed seine, there is a curtain net between the sink line and the bolch line. The bolch line is much longer than the foot line, and will be loose enough in fishing operation to get into the hole to scare out the fish which is hidden in the hole. The newly designed seine was tested in four Oreochromis nilotica pools in Yantai and two Oreochromis nilotica poo ls in Taian respect ively. The results show that the f ishing rate is over 60% for the first haul, and the total fishing rate for the f irst three hauls is above 90% . The newly designed seine is more suitable for the fishing operation in the pool with harder bottom.