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GH-IGF-I轴是鱼体体内一个重要的内分泌生理轴,主要调控鱼体的生长发育。用RT-PCR方法从加州鲈脑垂体和肝脏组织中分别扩增出加州鲈GH和IGF-I cDNA,克隆到pMD19 T-Vector上进行序列测定和分析。结果表明:1)加州鲈GH cDNA开放阅读框长为615 bp,编码204个氨基酸,其中信号肽17个氨基酸,成熟肽187个氨基酸。成熟肽中有四个保守的半胱氨酸残基(分别位于69,177,193,202),可形成两对二硫键。加州鲈GH氨基酸序列与蓝太阳鱼、斜带石斑鱼、金头鲷、虹鳟、鲤鱼、斑马鱼相比较,同源性分别为100%、97%、94%、66%、56%、53%。2)加州鲈IGF-I cDNA开放阅读框长为561bp,编码包括信号肽和B、C、A、D、E五个区域的186个氨基酸,形成成熟肽时,信号肽和E区域被切除。成熟蛋白的氨基酸序列与GenBank中已知的河鲈、三角鲂、金头鲷、舌齿鲈、斑马鱼的相比较,结果发现四个区域的保守性有差异,A区和B区保守性较高,C区和D区保守性较差。加州鲈GH和IGF-I cDNA的获得为进一步研究鱼体GH-IGF-I轴对生长发育的调控机制奠定了基础。 相似文献
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以勒氏笛鲷(Lutjanus russellii)基因组DNA为模板,采用同源克隆的方法,获得2 887 bp的肌肉生长抑制素(myostatin,MSTN)基因组序列,该MSTN序列具有3个外显子和2个内含子,包括101 bp的5′-UTR、385bp的外显子1、354 bp的内含子1、370 bp的外显子2、761 bp的内含子2、381 bp的外显子3和1 932 bp的3′-UTR。整个开放阅读框编码了378个氨基酸,前面的22个氨基酸为信号肽,具有9个保守的半胱氨基酸及一个RVRR的蛋白酶解加工位点。氨基酸序列分析发现,该基因编码的蛋白质与其他鱼类的I型同源性较高,与其他鱼类的2型MSTN同源性较低,且与鲈形目的同源性最高,与鲤形目的同源性较低,与人、鼠和鸡的同源性最低。采用邻接法(Neighbor-Joining)构建的MSTN的系统发育树表明,勒氏笛鲷MSTN与鲈形目鱼类的狼鲈属亲缘关系较近,且与鱼类的MSTN-1聚为1支。这表明该基因属于Ⅰ型MSTN基因。 相似文献
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草鱼呼肠孤病毒（Grass carp reovirus, GCRV）是草鱼出血病的病原.从患病草鱼体内分离到一株草鱼呼肠孤病毒GCRV 096,经反转录PCR,克隆得GCRV 096 长度为855 bp 的vp7 基因,并分析该基因编码蛋白的特性.结果表明,同一基因型分离株VP7 蛋白间的差异不大,但不同基因型分离株VP7 蛋白间存在很大的差异.对GCRV 096 VP7 蛋白生物信息学分析表明,信号肽位点位于20 氨基酸处,且VP7 含有4个潜在的抗原决定簇.构建GCRV 096 vp7 基因的酵母表达载体pGBKT7-S10,并成功转化至酵母中. 相似文献
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李亚 《广东海洋大学学报》2011,(4)
通过绿僵菌属甘露糖6-磷酸异构酶基因保守核苷酸区域设计简并性引物,采用RT-PCR及RACE-PCR技术成功克隆了金龟子绿僵菌mpi基因cDNA序列。该基因cDNA序列全长为1 513 bp,开放阅读框长度为1 328 bp,共编码441氨基酸。BLAST分析发现该基因演绎的氨基酸序列与其它真菌同源性较高,蛋白结构分析表明MPI蛋白是较保守的蛋白磷酸酶结构特征,主要由α螺旋和不规则卷曲构成。 相似文献
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应用已知的标签序列通过RACE-PCR和RT-PCR方法成功克隆红笛鲷(Lutjanus sanguineus)α-珠蛋白和β-珠蛋白基因的全长cDNA序列。α-珠蛋白和β-珠蛋白基因的cDNA全长分别为560和563 bp,开放阅读框分别为429和444 bp,各编码143和148个氨基酸,理论相对分子质量分别为15690和16400,理论等电点为7.79和6.61。氨基酸序列BLAST结果表明,红笛鲷α-珠蛋白基因和β-珠蛋白基因编码的氨基酸序列与其他鱼类的相似性分别在59%~79%和55%~80%之间。用邻接法(Neighbor-Joining)构建的α-珠蛋白基因和β-珠蛋白基因的系统发育树表明,红笛鲷与真鲷(Pagrus major)、金头鲷(Sparus aurata)和鰤(Seriola quinqueradiata)等亲缘关系较近。 相似文献
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April to October, 1997 comparative studios on the nitrogen budgets of closed shrimp polyculture systems showed that, in all the studied polyculture systems, nitrogen from feeds and fertilizers were the main input items, which comprised 70.7% - 83.9% of the total input nitrogen, 3.2% - 7.4% of which was provided by nitrogen fixation. It was in monoculture enclosures (Y-4, Y-11 and Y-12) that the percentage reached the maximum value. The output nitrogen in harvested products comprised 10.8% - 24.6% of total input nitrogen, and the highest percentage, 24.6%, was found in shrimp-fish-tagelus polyculture systems. In shrimp monoculture and shrimp-fish polyculture systems, they were 19.1% and 21.9%, respectively. The nitrogen utilization efficiency was different and varied from 12.2% to 20.1%. The highest, 20.1%, was found in shrimp-fish-tagelus polycultttre systems, and the average of 20.0% was found in shrimp-tagelus polyculture systems. The lowest, 12.2%, was found in shrimp monoculture systems.All the nitrogen utilization efficiencies in shrimp-fish systems or shrimp-scallop systems seemed to be higher than that of the monoculture system, but they showed little statistical difference. The main outputs of nitrngen were found in sediment mud, and comprised 48.2% -60.8% of the total input, the lowest percentage was found in shrimp-fish-tagelus polyculture systems, and the highost percentage in shrimp-scallop systems.During the experiment, nitrogen lost through denitrification and ammonia volatilization comprised 1.9% - 6.2%, averaged 2.8%, of the total input, and the loss through seepage comprised 5.9% - 8.9% of the total.The estimated nitrogen attached to the enclosure wall comprised 3.7% - 13.3% of the total, and was highest in shrimp monoculture systems. Compared with the classic shrimp farming industry, the closed shrimp polycul-ture systems may improve the nitrogen utilization efficiency, and hence reduce the environmental impacts on coastal waters. The nitrogen discharging rates for all the studied polyculture systems ranged from 3.0% to 6.0% of total input nitrogen. 相似文献
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Bothriocephalus acheilognathi is a potentially serious pathogen in wild or cultured fish in worldwide distribution. We examined 58-farmed grass carp from
Nanchang in the Changjiang (Yangtze) River drainage, from which 20.7% were found to harbor the parasite with an infection
intensity of 36.9±54.7. The parasites were identified based on morphology and rDNA ITS sequence analysis. The present report
represents the first record of the parasite in grass carp Ctenopharyngodon idella in the river drainage. 相似文献
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运用cDNA末端快速扩增（RACE）技术,克隆马氏珠母贝（Pinctadamαrtensii）热休克蛋白HSP60基因 cDNA全序列。序列全长2495坤,开放阅读框（ORF） 1734坤,编码577个氨基酸,预测的分子量约为61.79阳, 理论等电点为5.4905＇非翻译区（ 5＇UTR）长150坤, 3＇非翻译区（3＇UTR）长611 bp。同源性比对分析结果,马 氏珠母贝HSP60基因与与太平洋长牡妨（Crωsos仰a gIgω）和光滑双蹄螺（Biomphalaria glabrata）的同源性较 高,达到75%。氨基酸序列分析显示,马氏珠母贝HSP60氨基酸序列具有典型的rnt-HSP60特征序列、C-末端典型的GGM重复基序和一个ATP结合结构域。荧光定量数据分析发现,该基因在闭壳肌、外套腹、血淋巴、肝膜腺、性腺、躁、等6个组织中具有表达,在肝膜腺中表达量最高,腮中次之,而在血液和闭壳肌中仅有少量表达;在脂多糖剌激后,该基因表达水平上调,12 h后达到最大值,之后又逐渐下调,均高于对照组,差异具统计学意 义（P〈0.05）。 相似文献
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The inadequate nutrient content of pellet feeds widely used in recent years in China for grass carp farming led to lipid liver
degeneration in the fish. The present studies show that the pathological features of lipid liver disease are anaemia and hepatic
ceroidosis. Other clinical features are; the ratio of liver to body weight exceeds 3% and lipid content exceeds 5%. Extreme
infiltration of hepaiocytes by lipid results in the following deteriorative effects: swelling of the liver cells, increase
of lipid droplets in the cytoplasm and dislocation of the nucleus, loss of cytoplasm staining affinity, and increased activities
of GOT and GPT in serum. Lipid liver degeneration of grass carp can be divided into three stages: 1) deposition of liver lipid;
2) lipid infiltration of hepatic parenchyma; 3) atrophy of liver nucleus. The causes of lipid liver degeneration are complicated,
but the main cause is assumed to be an imbalance of nutrients in daily feed and the lock of some lipotropic substances.
A preliminary paper presented at the Asian Symposium on Freshwater Fish Culture, Beijing, October 11–15, 1985. 相似文献
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The intestinal bacteria of vertebrates form a close relationship with their host.External and internal conditions of the host,including its habitat,affect the intestinal bacterial community.Similarly,the intestinal bacterial community can,in turn,influence the host,particularly with respect to disease resistance.We compared the intestinal bacterial communities of grass carp that were collected from farm-ponds or a lake.We conducted denaturing gradient gel electrophoresis of amplified 16S rRNA genes,from which 66 different operational taxonomic units were identified.Using both the unweighted pair-group method with arithmetic means clustering and principal component analysis ordination,we found that the intestinal bacterial communities from the two groups of pond fish were clustered together and inset into the clusters of wild fish,except for DF-7,and there was no significant correlation between genetic diversity of grass carp and their intestinal bacterial communities(Mantel one-tailed test,R=0.157,P=0.175).Cetobacterium appeared more frequently in the intestine of grass carp collected from pond.A more thorough understanding of the role played by intestinal microbiota on fish health would be of considerable benefit to the aquaculture industry. 相似文献
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In this study, nest spawning was successfully induced by exogenous hormone injections and seawater flow stimulation, and optimum
condition for hatching fertilized eggs of burrow fish mudskipper,Boleophthalmus pectinirostris, was searched. Apart from spawning inside the nests, females also spawned outside the nests. The percentages of spawned nests
were 8.0% to 24.2%. Most eggs were observed adhered to the inner wall of the top half of the nest. Fertilization rates of
the nest-spawned eggs varied from 17.3% to 80.8%. Females could spawn after being artificially confined inside the nests with
males at ratios of 1∶1 or 1∶2, but the spawned eggs were not fertilized. Mean hatching rates of artificially fertilized eggs
incubated in round plastic buckets were 32.7%–70.6%, and in the net cages, were 4.2%–20.5%, respectively. Mean hatching rates
of nest-fertilized eggs incubated in the round plastic buckets were 33.6%–76.3%, and in the net cages, were 5.9%–25.2%. Results
showed that round bucket incubation was the best way for hatching fertilized eggs of mudskipper. Keeping the hatching seawater
flowing is an important way for increasing the hatching rates of the mudskipper fertilized eggs.
Supported by the NSFC (No. 30170739), and also supported by the Provincial Natural Science Foundation of Fujian Province (No.
B0210003). 相似文献
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用RT-PCR方法从1个H5N1亚型禽流感病毒分离株A/Chicken/Guangdong/DH/1997扩增NA基因cDNA片段,将其克隆至pMD18-T载体,获得重组质粒pMD-NA,并对其核苷酸序列进行测定和分析。结果表明,该毒株的NA基因长度为1350bp,编码449个氨基酸,与其它H5N1亚型AIV分离株的核苷酸序列同源性为97.0%～99.4%,氨基酸序列同源性为97.7%～99.1%,提示禽流感病毒NA基因保守性较高。NA基因氨基酸序列的聚类分析表明该毒株与来自香港的A/Pheasant/HK/FY155/01和A/Ch/HK/FY150/01两个分离株处于同一进化枝,亲缘关系较近。 相似文献
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五种鲤科鱼类生长激素cDNA的克隆和序列分析 总被引：1，自引：0，他引：1
通过RT-PCR方法,以草鱼、鳙鱼、鲫鱼、鲤鱼、齐口裂腹鱼等5种鲤科重要经济鱼类的垂体总RNA为模板扩增出其生长激素(fish growth hormone,fGH)的完整ORF序列,并克隆到pMD18-T载体上,命名为pMD-1(草鱼)、pMD-2(鳙鱼)、pMD-3(鲫鱼)、pMD-4(鲤鱼)、pMD-5(齐口裂腹鱼)。测序结果显示,ORF序列其长度均为633 bp。序列分析表明,所有ORF序列均以ATG为起始密码,以TAG为终止密码,编码210个氨基酸残基,推导的生长激素前体由22个氨基酸的信号肽和188个氨基酸的成熟肽组成。同源性分析表明,草鱼、鳙鱼、鲤鱼、鲫鱼和齐口裂腹鱼的fGH同源性在93.3%~99.5%之间。 相似文献