Studies on Pattern of Producing Transgenic Fish I.Production and Detection of All fish TRansgenic Common Carp With cMT sGH Gene

转基因鱼工作的开展被用于水产科学基础与应用研究的各个领域。应用方面的一项重要研究就是利用转生长激素基因来提高鱼的增长速度。需要解决两个问题：一是转基因鱼所使用的基因元件;二是外源基因的高整和率与高表达。已有许多报道认为：不同种动物的生长激素不一定相互促进生长。我们通过显微注射,将鲤鱼金属硫蛋白启动子（ｃＭＴ）与大马哈鱼生长激素基因（ｓＧＨ）的融合基因（ｃＭＴｓＧＨ）导入鲤鱼单细胞后期的早期胚胎,构建了全鱼转基因鲤鱼。通过斑点杂交、Ｓｏｕｔｈｅｒｎｂｌｏｔ结合ＰＣＲＳｏｕｔｈｅｒｎｂｌｏｔ,对外源ｓＧＨ的整和进行了精确的检测和分析。实验选取性成熟鲤鱼,收集卵子和精子,湿法受精获得受精卵。经过基因注射,孵化后的鱼苗放入水族箱。待鱼苗平游,提取总ＤＮＡ进行检测。以ＰｓｔＩ酶切质粒ｐｃＭＴｃＧＨ（Ｆｉｇ．４）分离３．４ＫｂｓＧＨ片段,用随机引物标记,作为杂交探针。同时,设计并合成ｓＧＨ基因的ＰＣＲ特异引物。Ｆｉｇ．１的斑点杂交结果与Ｆｉｇ．２的ＰＣＲＳｏｕｔｈｅｒｎ结果相比较（Ｔａｂｌｅ１）,说明斑点杂交存在着高的假阳性。而ＰＣＲＳｏｕｔｈｅｒｎ将ＰＣＲ的快速、方便与Ｓｏｕｔｈｅｒｎ的准确性相结合,排除了Ｐ     

鱼类转基因研究

本文综述了鱼类转基因研究的意义、鱼类基因转移研究的特点、转基因鱼的构建及检测技术、我国鱼类转基因研究的进展和鱼类基因转移研究中存在的主要问题。     

鱼类转基因研究

本文综述了鱼类转基因研究的意义、鱼类基因转移研究的特点、转基因鱼的构建及检测技术、我国鱼类转基因研究的进展和鱼类基因转移研究中存在的主要问题。     

猪生长激素基因在杆状病毒载体系统中的表达

通过对猪生长激素（ｐＧＨ）基因的ｃＤＮＡ进行测序,得到ｐＧＨｃＤＮＡ的全序列,并与Ｓｅｅｂｕｒｇ等报道的序列进行了比较和讨论。然后利用具人工合成启动子和多角体蛋白ＸＩＶ启动子的转移载体质粒ｐＳＸＩＶＶＩ＾＋Ｘ３／４构建出含ｐＧＨ基因的重组质粒ｐＸ３／４－ｐＧＨ。将ｐＸ３／４－ｐＧＨ与致死缺失型线性化ＡｃＭＮＰＶ－ＯＣＣ＾－ＤＮＡ共转染Ｓｆ９细胞,构建出既能形成多角体又能表达ｐＧＨ基因的苜蓿丫纹夜蛾     

泰山赤鳞鱼同工酶的研究

采用琼脂糖凝胶电泳或聚丙烯酰胺垂直板凝胶电泳（ＰＡＧＥ）技术研究了泰山赤鳞鱼早期发育阶段（受精后０—１２０ｈ）及成体眼、脑、心、肾、肝、肌６种组织中的ＬＤＨ同工酶分化表达谱式。结果表明：（１）赤鳞鱼在胚胎发育过程中Ｌｄｈ－Ａ基因和－Ｂ基因同时表达,形成５种不同形式的四聚体（Ｂ４、ＡＢ３、Ａ２Ｂ２、Ａ３Ｂ、Ａ４）。与大多数硬骨鱼相比,赤鳞鱼ＬＤＨ同工酶具有独特的早期个体发育谱式：在整个胚胎发育时期,Ａ亚基与Ｂ亚基的活性几乎相等。（２）赤鳞鱼的ＬＤＨ同工酶谱具有明显的组织特异性。Ｌｄｈ－Ｃ基因仅在肝脏组织表达,以向阴极迁移的分子形式（ＬＤＨ－Ｃ４）特异地表达。     

GHcDNA克隆在大肠杆菌中表达将有望应用于水生鱼类动物

猪生长激素 (ＧＨ)为单一的多肽 ,无亚基 ,不要糖基化修饰 ,只含 2个硫键 ,有重复的特性 ,并能在大肠杆菌体内进行表达 ,其表达量为 2 0 .5 %。但在构建鲑鱼ＧＨ、鲤鱼ＧＨ的表达载体分别为 ｐＢＶｓＧＨＩ8和ｐＢ ＶｃＧＨ8时 ,其表达量分别是 10 %和 2 9.2 %。当猪ＧＨ基因同用大肠杆菌、鲑、鲤进行表达 ,其表达载体均为ｐＢＶ2 0 0时 ,各个表达量仍有很大差异。这主要是外源基因在同一表达载体中所表达的水平不同有关 ,亦即外源基因 5′端ＡＵＧ附近的ＲＮＡ二级能量的大小、ＡＵＧ与ＳＤ序列之间的距离大小 ,外源基因 3′测翼稳定性的强…     

生长激素及其基因转移对鱼类生长和渗透压的调节作用

生长激素及其基因转移对鱼类生长和渗透压的调节起着重要作用。转生长激素基因鱼表现出明显的快速生长效应。生长激素促进了鲑科鱼类对海水的适应能力。本文对此进行了综述,并讨论了生长激素及其基因转移对鱼类生长和渗透压调节作用的生理机制、生长激素与胰岛素样生长因子以及甲状腺激素的关系。     

生长激素及其基因转移对鱼类生长和

生长激素及其基因转移对鱼类生长和渗透压的调节起着重要作用.转生长激素基因鱼表现出明显的快速生长效应.生长激素促进了鲑科鱼类对海水的适应能力.本文对此进行了综述,并讨论了生长激素及其基因转移对鱼类生长和渗透压调节作用的生理机制、生长激素与胰岛素样生长因子以及甲状腺激素的关系.     

总DNA介导鱼类基因转移的初步研究

自七十年代我国开始将外源总DNA转移技术应用于棉花和水稻品种改良的研究中,但至今尚未见有关鱼类总DNA转移成功的报道。本实验采用显微注射和精于载体的方法将鲫鱼肝总DNA转移到红鲤受精卵,利用鯽鱼和红鲤在孵化前后色素表达的差异来筛选转移基因个体。此方法无需克隆目的基因及DNA重组操作,而且筛选简便,目的在于探讨总DNA转移方法应用于鱼类品种改良的可能性。     

野生型及启动子区突变的HPFHAγ基因在MEL细胞中的表达

研究了野生型Ａγ珠蛋白基因、中国型胎儿血红蛋白持续存在综合症（ＨＰＦＨ）Ａγ基因（启动子区─１９６Ｃ→Ｔ突变）及希腊型ＨＰＦＨＡγ基因（─１１７Ｇ→Ａ突变）在鼠红白血病（ＭＥＬ）ＧＭ９７９细胞中的表达。比较每个整合的Ａγ基因表达的ｍＲＮＡ量,─１９６Ｃ→Ｔ突变Ａγ基因未显示比野生型Ａγ基因明显增加的表达水平,而─１１７Ｇ→Ａ突变Ａγ基因在未分化的及ＨＭＢＡ或丁酸钠诱导分化的ＭＥＬＧＭ９７９细胞中的表达水平增加到野生型Ａγ基因相应水平的２．３倍至３．１倍。此结果提供了又一个实验证据,说明启动子─１１７Ｇ→Ａ突变是希腊型ＨＰＦＨ中胎儿Ａγ基因在成人中持续活跃表达的原因,这也为β地中海贫血和镰刀性贫血的基因治疗提供了新途径,即可以考虑用转移─１１７Ｇ→Ａ突变Ａγ基因的方法改善患者的贫血症状。     

Fish cell lines: Two new cell lines derived from explants of trunk musculature ofCynoscion arenarius

Summary Two cell lines were established from explants of trunk musculature of healthy, males sand seatrout,Cynoscion arenarius. One of the lines, designated CyA-1, has been carried through 150 subcultures during 6 yr. The other, designated CyA-2, has been carried through 100 subcultures during 2 yr. Both lines grow well in L15 medium adjusted to 0.150M NaCl and supplemented with 10% fetal bovine serum. Optimal growth occurs at temperatures between 24 and 30°C. The species of origin of both lines was confirmed by a cytotoxic antibody dye exclusion test. The karyotype of CyA-1 has not yet stabilized, showing a modal chromosome number of 120 at Passage 9, 89 at Passage 63, and 79 at Passage 100. The karyotype of CyA-2 is rather stable, with a modal chromosome number of 47 at Passage 1 and 49 at Passage 100. Chromosome morphology of CyA-2 is homogeneous (small, acrocentric), whereas the chromosomes of CyA-1 show considerable size variation (with small chromosomes possibly formed from fragmentation of original structures). Both lines were found to be free of bacterial or fungal contamination. Both lines supported replication of lymphocystis virus strains isolated fromBairdiella chrysura (the silver perch) and fromMicropogon undulatus (the Atlantic croaker) but were refractory to 11 other viruses (4 from fish, 1 from amphibians, and 6 from mammals). This study was supported in part by the National Oceanographic and Atmospheric Administration, Office of Sea Grant No. 04-3-158-58.     

Histopathology, blood chemistry, and physiological status of normal and moribund striped bass (Morone saxatilis) involved in summer mortality ('die-off') in the Sacramento-San Joaquin Delta of California

Summer mortality ('die-off') is common in striped bass, Morone saxatilis (Walbaum), in the San Francisco Bay-Delta region. Tissue and blood samples of moribund and healthy striped bass collected during the summers of 1986–1988 were analysed. Sixteen moribund and 25 healthy reference fish from the Carquinez Strait area and eight fish caught in the Pacific Ocean were studied. Moribund fish plasma was invariably yellow-orange; most of the moribund fish had discoloured livers with haemorrhagic regions, and approximately one-third had haemorrhagic intestines. Piasma levels of aspartale aminotransferase, uric acid, alkaline phosphatase and cortisol were significantly higher than in reference fish from Carquinez Strait and the Pacific Ocean, whereas cholesterol, sodium, chloride, triiodothyronine and glucose levels were significantly lower. Hepatic heavy metal concentrations and bacterial content were similar in moribund and reference fish. Gill Na.⁺, K⁺ -ATPase activity was significantly lower in moribund fish. Liver, kidney, intestine, and thyroid follicles of moribund fish displayed various histopathological changes, and corticosteroidogenic (interrenal) tissue could not be identified positively in moribund fish. These findings are discussed in relation to recent work on the chemical burdens (industrial and agricultural hydrocarbons) found in livers from some of the fish examined in this study.     

南海东沙岛及太平岛鱼类种类组成和动物地理学特点

就笔者等在南海之东沙岛及太平岛两处海域所调查记录到的珊瑚礁鱼类,连同自1975年以来之文献与数据库的记录作综合整理.结果表明,东沙岛有73科652种,太平岛有56科466种,两岛合计共有76科759种.太平岛所记录的总鱼种数较东沙岛为少,主要原因纯系因太平岛之礁域面积较小,且调查次数甚少之故.根据各鱼种地理分布范围之分析,发现在此二岛所记录到的鱼种几乎全部都是印度-太平洋、印度-西太平洋及西太平洋等广泛分布的鱼种,目前只有黄头刻齿雀鲷(Chrysiptera chrysocephala)1种是只在南海发现之鱼种.由此可推测南海的鱼类相应与周边海域或珊瑚三角的鱼类相充分交流,并未有明显隔离或种化的现象.东沙岛及太平岛鱼类相之Sφrensen相似性系数值虽已达85.69％,但其中仍有240种是只分布在东沙岛及台湾(165种),或太平岛及台湾(75种),而未在另一岛屿发现之鱼种,且其中只在太平岛及东沙岛发现的21种及20种鱼中,大多为稀有种,非常不容易被调查到.故推测目前在此二岛所作的调查频度相当不足,如果调查能更密集更长期,则两岛的鱼相会更相似.     

海水鱼类共附生细菌群落研究进展

与海水鱼类处于共生、共栖、寄生或附生关系的细菌群落,称之为海水鱼类共附生细菌群落。这类细菌群落生活在海水鱼体表(皮肤、鳃)以及体内(消化道和血液、肌肉、肝脏、肾脏等内部组织器官),它们彼此之间以及与宿主之间存在着极其密切的关系,并且对于宿主的健康生长具有重要作用。然而,由于目前对养殖系统中的微生物群落中的致病微生物缺乏有效控制措施,处于迅速发展中的养殖产业经受着相当严重的疾病问题。因而,生态健康养殖被提到议事日程上来,并且日益受到重视,其中微生物生态调控是极其重要的一个环节。由于目前相关的基础微生物生态学资料比较缺乏,尤其是在国内作为微生物生态的基本组成部分,与微生物病害发生有着直接关系的大多数海水养殖鱼类的共附生细菌群落研究资料相当缺乏。因此,很有必要开展海水养殖鱼类共附生细菌群落相关研究。在此背景下,从研究的目的和意义以及国内外研究现状包括宿主海水鱼类种类、共附生细菌群落类别、共附生细菌群落的影响因素、共附生细菌群落对宿主的作用等方面综述了海水鱼类共附生细菌群落的研究进展,并对此类研究趋势进行了展望,为开展相关研究提供一定的参考。     

Biological Inspiration: From Carangiform Fish to Multi-Joint Robotic Fish

This paper presents a novel approach to modelling carangiform fish-like swimming motion for multi-joint robotic fish so that they can obtain fish-like behaviours and mimic the body motion of carangiform fish. A given body motion function of fish swimming is firstly converted to a tail motion function which describes the tail motion relative to the head. Then, the tail motion function is discretized into a series of tail postures over time. Thirdly, a digital approximation method calculates the taming angles of joints in the tail to approximate each tail posture; and finally, these angles are grouped into a look-up table, or re-gressed to a time-dependent function, for practically controlling the tail motors in a multi-joint robotic fish. The paper made three contributions: tail motion relative to the head, an error function for digital approximation and regressing a look-up table for online optimization. To prove the feasibility of the proposed methodology, two basic swimming motion patterns, cruise straight and C-shape sharp turning, are modelled and implemented in our robotic fish. The experimental results show that the relative tail motion and the approximation error function are good choices and the proposed method is feasible.     

Karyotypic diversity of the armoured catfish genus Hypostomus (Siluriformes: Loricariidae) in the context of its occurrence and distribution

This study taxonomically reviewed the specimens studied by Artoni & Bertollo (1996) and assimilated species of Hypostomus into three groups according to their cytogenetic characteristics, vagility and occurrence environments.     

三倍体湘云鲫繁殖季节的性腺结构观察

在繁殖季节解剖３７尾龄和１６尾声龄湘云鲫,用光镜和电镜观察了湘云鲫的性腺结构。在湘鲫的性腺部位有三种类型的结构：（１）清巢型;精巢小叶内分布有精子细胞,精子细胞呈退化现象,没有发现有正常成熟精子。（２）卵巢型,该类型性腺的大部分为体积较小,分化程度较低的呈囊状排列的细胞,在这些小细胞中镶嵌着少量初级卵母细胞和体积较大的退化的卵母细胞。（３）脂肪型;该类型的性腺部位仅有两条脂肪组织,即没有卵巢结构,     

我国转基因鱼研制的历史回顾与展望

中国自从诞生了首例转基因鱼以来,在后续30多年里取得了一系列重要研究进展。全球范围的转基因鱼研究包括多种养殖鱼类,目标性状涉及快速生长、抗病抗逆和品质改良。现在已经初步建成转基因鱼育种技术体系和安全评估体系,为转基因鱼产业化奠定了重要基础。本文以转基因黄河鲤育种研究为主线,简要回顾了转基因鱼研究的发展历程,并对转基因鱼育种面临的问题和发展前景进行了分析和展望。     

Fish parasitism in a new impoundment and differences upstream and downstream

The parasitic fish fauna in Guelph Lake, a four-year old impoundment, were compared with those in upstream and downstream localities. The lacustrine assemblage of the young mesotrophic to eutrophic impoundment was dominated by the cestodes, Ligula intestinalis and Proteocephalus ambloplites, and the nematode, Philometra sp. The acanthocephalan, Neoechinorhynchus saginatus, and the nematode, Rhabdacona cascadilla dominated the rheophilous assemblage of fish parasites, but the prevalence rate and intensity of R. cascadilla was significantly reduced downstream.     

Habitat use of the European mudminnow Umbra krameri and association with other fish species in a disconnected Danube side arm

Fish assemblages along the longitudinal course of an old, disconnected and modified side arm of the Danube floodplain downstream of Vienna, Austria, as well as habitat structure, hydro‐morphological and hydro‐chemical factors, were investigated in order to analyse the key environmental determinants of the European mudminnow Umbra krameri. Generally, U. krameri was the most abundant species in the system. It occurred in disconnected ditches, ponds and pools with dense reed belts and comparatively low nutrient content, indicating its natural association with marsh habitats. At infrequently disturbed sites it was associated with a small group of stagnophilious and highly specialized species with adaptations to strong oxygen fluctuations. At frequently flooded sites, the species was absent or occurred in low abundances, indicating its adaptation to water bodies in older successional stages and its low competitive power in permanently connected floodplain habitats.