日本海马仔鱼消化系统的组织学研究

日本海马仔鱼消化系统的组织学研究ＡＳＴＵＤＹＯＮＴＨＥＨＩＳＴＯＬＯＧＹＯＦＤＩＧＥＳＴＩＶＥＳＹＳＴＥＭＯＦＬＡＲＶＡＬＳＥＡ－ＨＯＲＳＥ张峰（大连水产学院，１１６０２３）ＺｈａｎｇＦｅｎｇ（ＤａｌｉａｎＦｉｓｈｅｒｉｅｓＣｏｌｅｇｅ，１１６０２３...     

南海近底层拖网的设计和试捕效果

南海近底层拖网的设计和试捕效果ＤＥＳＩＧＮＡＮＤＦＩＳＨＩＮＧＥＸＰＥＲＩＭＥＮＴＡＬＲＥＳＵＬＴＯＦＮＥＡＲ－ＢＯＴＴＯＭＴＲＡＷＬＩＮＳＯＵＴＨＣＨＩＮＡＳＥＡ杨吝（中国水产科学研究院南海水产研究所，广州５１０３００）ＹａｎｇＬｉｎ（ＳｏｕｔｈＣ...     

氨对草鱼生长的危害

氨对草鱼生长的危害朱耘，吴圣杰，华丹（中国水产科学研究院淡水渔业研究中心，无锡２１４０８１）关键词氨，草鱼，生长ＴＨＥＨＡＲＭＦＵＬＥＦＦＥＣＴＯＦＴＨＥＡＭＭＯＮＩＡＯＮＴＨＥＧＲＯＷＴＨＯＦＧＲＡＳＳＣＡＲＰ（ＣＴＥＮＯＰＨＡＲＹＮＧＯＤＯＮＩＤ...     

不同脂肪源饲料对草鱼稚鱼生长的影响

不同脂肪源饲料对草鱼稚鱼生长的影响刘玮，徐萍，任本根，龚纲明（江西省科学院生物资源研究所，南昌３３００２９）关键词草鱼，鱼饲料，必采脂肪酸ＥＦＦＥＣＴＳＯＦＤＩＥＴＳＣＯＮＴＡＩＮＩＮＧＤＩＦＦＥＲＥＮＴＬＩＰＩＤＳＯＮＧＲＯＷＴＨＯＦＪＵＶＥＮＩＬ...     

罗非鱼饲料中海带渣含量对氨基酸表观消化率的影响↑（*）

罗非鱼饲料中海带渣含量对氨基酸表观消化率的影响ＥＦＦＥＣＴＯＦＬＡＭＩＮＡＲＩＡＲＥＳＩＤＵＥＳＯＮＡＰＰＡＲＥＮＴＤＩＧＥＳＴＩＢＩＬＩＴＹＯＦＡＭＩＮＯＡＣＩＤＳＩＮＤＩＥＴＯＦＯＲＥＯＣＨＲＯＭＩＳＮＩＬＯＴＩＣＵＳ甘纯玑彭时尧施木田（福建农业...     

用酶联免疫吸附受体法检测鱼类生长激素的生物活性

根据激素一受体反应的原理，结合酶联免疫吸附测定法（ＥＬＩＳＡ）和放射受体测定法（ＲＲＡ）的优点，应用我们纯化的Ａ昌鱼生长激素（ｇｃＧＨ）和大鳞大马哈鱼生长激素（ｓＧＨ）及其特异抗体，采用鱼类肝细胞膜受体制剂，首次建立了测定鱼类ＧＨ生物活性的酶联免疫吸附受体测定法（ＥＬＩＳＡ－ＲＡ）。此法检测草鱼ＧＨ的灵敏度达０．０６３￣０．１２５ｕｇ／ｍｌ，检测大马哈鱼ＧＨ与草鱼肝膜受体结合的灵敏度达０．２５ｕｇ     

草鱼Mx蛋白基因的克隆与原核表达

根据已报道的Mx蛋白cDNA序列设计合成特异引物，应用逆转录-聚合酶链式反应(RT-PCR)方法，从草鱼呼肠孤病毒(GCRV)诱导的草鱼(Ctenopharyngodon idellus)肝脏总RNA中扩增获得Mx蛋白cDNA，回收纯化后克隆到pGEM-T Easy Vector系统的T载体上。重组子的序列分析表明：所克隆的Mx蛋白cDNA长722bp，编码240个氨基酸，包括1个三联GTP结合区域和1个发动蛋白(dynamin)族特征的序列。与已知鱼类Mx蛋白基因序列比较表明，草鱼Mx蛋白基因序列与其它鱼类Mx基因相应序列具有较高的同源性，其中与斑马鱼Mx蛋白E型基因碱基序列比较同源性最高，为87．1％。将此基因改造后定向克隆至原核表达质粒pBV220，构建成重组草鱼Mx蛋白基因表达质粒pBVgcMx，并在大肠杆菌中获得高效表达，重组草鱼Mx蛋白的表达量占菌体总蛋白的26．6％。Q-sephrose FF层析柱分离纯化的重组草鱼Mx蛋白纯度达95．6％。     

鱼类仔鱼期的摄食和生长

鱼类仔鱼期的摄食和生长殷名称（上海水产大学，２０００９０）关键词鱼类，仔鱼期，摄食，生长ＦＥＥＤＩＮＧＡＮＤＧＲＯＷＴＨＯＦＴＨＥＬＡＲＶＡＳＴＡＧＥ．ＯＦＦＩＳＨ￥ＹｉｎＭｉｎｇｃｈｅｎｇ（ＳｈａｎｇｈａｉＦｉｓｈｅｒｉｅｓＵｎｉｖｅｒｓｉｔｙ，２...     

INGESTIONANDEXCRETIONOFSEVERALANTIBACTERIALDRUGSBYTHEBRINESHRIMP（ARTEMIAPARTHENOGENETICA）1．TOXICITYT

ＩＮＧＥＳＴＩＯＮＡＮＤＥＸＣＲＥＴＩＯＮＯＦＳＥＶＥＲＡＬＡＮＴＩＢＡＣＴＥＲＩＡＬＤＲＵＧＳＢＹＴＨＥＢＲＩＮＥＳＨＲＩＭＰ（ＡＲＴＥＭＩＡＰＡＲＴＨＥＮＯＧＥＮＥＴＩＣＡ）１．ＴＯＸＩＣＩＴＹＴＯＡＲＴＥＭＩＡ，ＩＮＧＥＳＴＩＯＮＡＮＤＥＸＣＲ...     

淡水鱼类急性中毒死亡的判断

淡水鱼类急性中毒死亡的判断ＤＩＡＧＮＯＳＩＳＦＯＲＦＲＥＳＨＷＡＴＥＲＦＩＳＨＫＩＬＬＳＣＡＵＳＥＤＢＹＡＣＵＴＥＰＯＩＳＯＮＩＮＧ曹立业李应仁（中国水产科学研究院，北京１０００３９）ＣａｏＬｉｙｅＬｉＹｉｎｇｒｅｎ（ＣｈｉｎｅｓｅＡｃａｄｅｍｙｏｆ...     

哲罗鲑胰岛素样生长因子－I（IGF－I）cDNA 分子克隆、序列分析及组织表达

采用逆转录聚合酶链式反应（RT－PCR）方法,从哲罗鲑（Hucho taimen）肝脏的总 RNA 中扩增出胰岛素样生长因子－I（IGF－I）的 cDNA 开放阅读框（Open reading frame, ORF）序列,运用软件对其进行生物信息学分析,并利用荧光实时定量 PCR 技术检测了哲罗鲑成鱼不同组织中 IGF－I mRNA 的表达情况。结果显示, IGF－I 基因的 cD－NA 开放阅读框为573 bp,编码190个氨基酸,蛋白质等电点为9．21,氨基酸结构由信号肽、 B、 C、 A、 D 结构域及 E 肽组成;氨基酸序列与其他鲑科鱼类具有较高的同源性,其中与北极红点鲑的 IGF－I 同源性最高（99．2％）;组织表达分析显示,哲罗鲑 IGF－I mRNA 在肝脏中表达量最高,在鳃、前肠中次之,在脑、头肾、脾、心、胃和肌肉等组织中的表达量较低。     

Cloning, tissue distribution and hormonal regulation of stearoyl-CoA desaturase in tilapia, Oreochromis mossambicus

The stearoyl-CoA desaturase cDNA in tilapia (Oreochromis mossambicus) was cloned by RT-PCR and RACE, and it was compared with those in grass carp, common carp and milkfish. Nucleotide sequence analysis revealed that the full length of cDNA (1172 bp) clone encompasses 1008 bp open reading frame (ORF) encoding 336 amino acid residues. The deduced amino acid sequence shares 78–82% identity with the teleosts and 64–66% with mammals compared, and like these fish, the cloned tilapia stearoyl-CoA desaturase amino acid sequence conserves three histidine cluster motifs (one HXXXXH and two HXXHH), which functioned as non-heme iron binding sites, essential for stearoyl-CoA desaturase activity. RT-PCR and Northern blot analysis reveal that tilapia stearoyl-CoA desaturase is expressed only in liver, but the stearoyl-CoA desaturase expression in multiple tissues was observed in milkfish, grass carp and carp. Further, the hormonal regulation of stearoyl-CoA desaturase gene expression was investigated by a single injection of 17β-estradiol and testosterone. The results showed that the administration of 17β-estradiol to tilapia led to a greater increase in desaturase activity than testosterone, and higher doses of steroids produced greater increases in enzyme activity. The comparative RT-PCR analysis showed that the stearoyl-CoA desaturase mRNA level increased significantly in 17β-estradiol treated animals, especially in the groups receiving a single injection of 50 mg 17β-estradiol. This was reflected in the decrease in the saturated fatty acids and the increase in the monounsaturated fatty acids. The proportion of the polyunsaturated fatty acids was not affected.     

Occurrence of two distinct molecular species of cathepsin B in carp Cyprinus carpio

ABSTRACT:   We purified cathepsins B₁ and B₂ from the ordinary muscle of carp Cyprinus carpio . The N-terminal amino acid sequences (12 residues) of 29 kDa bands of cathepsins B₁ and B₂ are the same and showed high homology of 75% and 83%, respectively, with the heavy chain of rat and human cathepsins B. Based on conserved sequences of other cathepsins B and the N-terminal amino acid sequences of 29 kDa bands, we cloned carp cathepsin B cDNA. The nucleotide sequence of carp cathepsin B cDNA consists of 1470 bp including a 993 bp open reading frame, encoding a deduced protein of 330 amino acids. The deduced amino acid sequence of carp cathepsin B has similarity of 80% to rainbow trout cathepsin B and of 76–78% to other vertebrate cathepsins B. The sequence of its isoform was also determined during molecular cloning, which has 94.8% similarity with first cloned cathepsin B. They are completely same in N-terminal amino acid sequence of heavy chain, active site and potential N-glycosylation site. This indicates there are at least two kinds of cathepsin B functioning in vivo in carp.     

Molecular cloning of Insulin-like Growth Factor-I from triangular bream (Megalobrama terminalis)

Triangular bream (Megalobrama terminalis) IGF-I DNA and gene were cloned from triangular bream liver for the first time by RT-PCR. Sequence analysis indicated that the IGF-I cDNA consisted of 486 nucleotides encoding 117 amino acids which spanned the complete signal peptide, B, C, A, D and E domains. Analysis of the E domain indicated that triangular bream IGF-I belongs to the IGF-I Ea-2 subtype. Compared to bluntnose bream (Megalobrama amblycephala), another member of the same Megalobrama genus, triangular bream IGF-I shared 99.8% identity in cDNA sequence and 99.4% in predicted amino acid sequence. However, considerable differences were found in comparison to with common carp (Cyprinus carpio) and grass carp (Ctenopharyngodon idellus), which are members of the same family but of a different genus. This revised version was published online in July 2006 with corrections to the Cover Date.     

草鱼胞质苹果酸脱氢酶(cMDH)的序列克隆及分析

以实验室构建的草鱼(Ctenopharyngodon idellus)肠道cDNA文库苹果酸脱氢酶(cMDH)EST序列为基础,采用末端快速扩增法(RACE),从健康雌性草鱼(Ctenopharyngodon idellus)肠道细胞RNA中获得1391 bp的草鱼肠道cMDH的cDNA序列(NCBI登录号:EU569765)。结果显示:该序列包含62 bp的5′非编码区(5′-UTR),330 bp的3′非编码区(3′-UTR),典型加尾信号AATAA位于polyA起点上游16 bp。开放阅读框ORF长999 bp,共编码333个氨基酸,预测蛋白等电点为6.67,大小为36 kDa。多序列比对显示草鱼胞质苹果酸脱氢酶具有典型的苹果酸脱氢酶功能区保守序列,该酶与斑马鱼cMDH相似度最高达到95.5%,与模式植物拟兰介的相似度为57.8%,其预测三级结构具有典型cMDH功能区。Southern杂交结果表明草鱼cMDH属于多拷贝基因家族。     

草鱼SREBP-1基因的克隆及糖对其在肝脏中表达的影响

固醇调节元件结合蛋白(SREBPs)是调控糖脂代谢相关基因表达的关键核转录因子。为获知草鱼SREBP-1基因的序列及其在肝脏中的表达规律,本实验采用同源克隆和RACE方法获得了草鱼SREBP-1基因的部分cDNA序列,并通过生物信息学方法对该基因及所编码蛋白的结构特征进行了分析;采用实时荧光定量PCR技术,对SREBP-1基因在8种不同组织的表达规律及低糖(糖含量24%)和高糖(糖含量42%)投喂条件下肝脏中的表达水平进行了研究。结果显示,所克隆到的草鱼SREBP-1基因cDNA长4 760 bp,其中包括开放读码框3 426bp,编码1 141个氨基酸;草鱼SREBP-1具有一个典型的碱性螺旋-环-螺旋亮氨酸拉链结构(bHLH-zip);氨基酸序列比对结果显示,草鱼SREBP-1与其他鱼类的同源性在76%~88%之间,与斑马鱼的进化关系最近;草鱼SREBP-1基因在脑中的表达量最高,肝脏和肠次之,在肾脏、脾脏、肌肉、脂肪和性腺中均有少量表达;与对照组相比,SREBP-1基因在高糖诱导下表达量显著提高(P0.05),低糖诱导下没有显著性差异。研究表明,在高糖负荷条件下,草鱼肝脏中SREBP-1可能会促进糖的利用和转化,从而参与糖代谢调节过程,为丰富鱼类糖代谢调控机理提供研究资料,并有望为提高鱼类对饲料糖的利用效率提供理论依据。     

长江草鱼群体MHC Class ⅡB的多态性和进化分析

克隆及测序草鱼(Ctenopharyngodon idella)长江3个群体的主要组织相容性复合体(MHC)Class II B基因编码β1和β2区的第2和第3个外显子及两个外显子之间的内含子,分析了草鱼MHC的进化模式和种群遗传结构。结果显示:实验共定义了34个等位基因,每条序列包括长为130~136 bp的第2个外显子,长为218 bp的第3个外显子以及长81~371 bp的内含子。序列分析揭示,第2个外显子有106个核苷酸变异位点(78%)和40个氨基酸变异位点(88%),而第3个外显子有100个核苷酸变异位点(45%)和41个氨基酸变异位点(56%),β1变异要大于β2区。用β1和β2区序列分别构建的邻接(NJ)系统树均显示5个具有高支持率的谱系,结合序列变异特点和内含子长度,推测草鱼至少存在5个MHC Class II座位。分别计算β1的肽结合位点(PBR)、非肽结合位点及β2的非同义替换率(dN)和同义替换率(dS),PBR的dN/dS为2.03(P<0.05),非肽结合位点和β2则小于1,表明草鱼MHC受到歧化选择作用。根据等位基因在群体中的分布频率作分子方差分析(AMOVA),得出FST为0.37%,提示长江草鱼MHC没有遗传分化。     

Cloning, characterization and expression of the LECT2 gene in grass carp

An expressed sequence tag of grass carp leukocyte cell–derived chemotaxin 2 (LECT2) gene was screened from an established intestinal cDNA library. Rapid amplification of cDNA ends gave rise to a full-length LECT2 cDNA (gcLECT2) with a complete open-reading frame of 474 bp, encoding 158 amino acids about 17.9 kDa. Homology search and sequence alignment showed that this deduced protein sequence shared a high identity with LECT2 from other vertebrates. Western blotting indicated immunological cross-reactivity occurs between grass carp and human LECT2 protein. This gcLECT2 genomic sequence is 1,868 bp in size, which consists of five exons and four introns. Real-time quantitative PCR analysis revealed that gcLECT2 gene is ubiquitously expressed in different tissues of healthy grass carp including brain, gut, liver, spleen, kidney, muscle and heart, while the expression levels were significantly increased in liver and spleen followed by Aeromonas salmonicida infection. 992 bp 5′-flanking region sequence was cloned and analyzed, where one CAAT box and one GC island were found. Our results showed that the LECT2 is suggested to be most possibly involved in the grass carp’s immune response.