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1.
采用聚合酶链反应(PCR)方法改造鲑鱼(Oncorhynchustschawytscha)生长激素cDNA。改造后的cDNA去除了编码生长激素成熟肽cDNA以外的信号肽序列及5′端和3′端的非翻译序列,加入一起始密码,并在改造后的基因两端引入BamHI识别序列各1个。亚克隆后对改造过的基因进行了序列分析。  相似文献   

2.
用酶联免疫吸附受体法检测鱼类生长激素的生物活性   总被引:13,自引:0,他引:13  
陈松林 《水产学报》1995,19(3):217-224
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  相似文献   

3.
草鱼血清生长激素水平的日变化   总被引:1,自引:0,他引:1  
张为民 《水产学报》1995,19(3):263-267
草鱼血清生长激素水平的日变化DAILYVARIATIONSOFSERUMGROWTHHORMONELEVELSINGRASSCARP¥ZhangWeiminandLinHaoran(BiologyDepartmeatofZhongshanUniver...  相似文献   

4.
将大鳞大麻哈鱼的垂体细胞特异性表达的生长激素基因的启动子与白喉毒素基因的A链相连,构建成融合基因GH-DTA。通过显微注射导入金鱼受精卵内,获得的鱼经Southern blot分子杂交检测表明,少数实验鱼染色体DNA中含有导入的外源基因。胚胎的原位分子杂交和垂体的组织切片观察发现,转基因鱼和正常鱼之间存在差异。  相似文献   

5.
1注重亲鱼的选择选择的雌鱼要求腹部膨大、松软 ,卵巢轮廓明显 ,外生殖乳突出而圆 ;雄鱼外生殖乳突长而尖 ,胸鳍刺上有强大锯齿。2适时催产选择性成熟的亲鱼在3~4月产卵盛期 ,天气比较稳定、水温适宜(20~23℃)的时间进行催产。3催产是关键笔者对4龄大口鲶共25尾催产全部获得成功。雌鱼17尾 ,总重50.2kg;雄鱼8尾 ,总重16.5kg。催产水温为23~23.5℃ ,选用宁波激素制品厂生产的HCG和LRH A作催产剂。3.1注射剂量雌鱼第1针剂量为HCG1000IU/kgHCG LRH A1.5μg/kg 。…  相似文献   

6.
鱼类基因分子生物学及转基因鱼研究进展↑(*)   总被引:4,自引:0,他引:4  
真核基因分子克隆技术的建立,提供了从核苷酸水平了解鱼类基因结构的手段。鱼类基因的分子克隆始于八十年代初、中期对虹鳟(Salmogairdneri)鱼精蛋白基因[52]和美洲黄盖鲽(Psendopleuronectesamericanus)抗冻蛋白(AFP)基因[22]的分子克隆,随后扩展到其他极地鱼类AFP基因,接着扩展到鱼类生长激素(GH)基因、催乳激素(PRL)基因、生长催乳素(somatolactinSL)基因等数十个基因。对这些控制鱼类重要生理机能的基因结构的认识,无论在理论上还是在渔业…  相似文献   

7.
条纹石鮨摄食强度与代谢及能量收支间的关系   总被引:1,自引:0,他引:1  
设日摄食率为0.4%、1.5%、4.0%三种水平,分别测定条纹石鱼旨Moronesaxatilis幼鱼的代谢及能量的收支情况。处于饱食(日摄食率4.0%)及维持状态(日摄食率1.5%)下,该鱼代谢率增加的峰值分别于摄食后3h及7h内出现,为静止时代谢水平的2.10倍和2.46倍,SDA(特殊动力作用)持续时间均为21h,SDA总耗能量分别占摄入能的13%和35.8%。条纹石鱼旨在饱食时的转化效率K(湿重)为20.85%。三种摄食水平下条纹石鱼旨的能量收支方程分别为:饱食状态(日摄食率4.0%)时,100C=8.4F+7.8U+13.0SDA+36.9(Rs+Ra)+33.9G;维持状态(日摄食率1.5%)时,100C=7.8F+4.1U+35.8SDA+52.3(Rs+Ra),其中G=0;减重状态(日摄食率0.4%)时,(F+U+R)/(C+P1)=67.3%,其中C+P1=14.4C+85.6P1,F+U+R=1.8F+8.1U+57.4R。  相似文献   

8.
WSSV 3个编码蛋白的基因密码子偏爱性分析   总被引:4,自引:0,他引:4  
运用EMBOSS(The European Molecular Biology Open Software Suite)的CHIPS(Condon Heterozygosity in a Protein Coding Sequence)和CUSP(Create a condon usage table)程序对WSSV3个编码蛋白的基因密码子偏爱性分析,分析结果与大肠杆菌、酵母及人的密码子偏爱性进行比较。结果显示,WSSV—vp37、WSSV—vp39及WSSV—vp19的Nc(Effective number of condons)值为49.897、48.640、46.868。3种蛋白的编码密码子使用频率有较大的差异,其在大肠杆菌、酵母及人的密码子偏爱性也有不同,分析结果表明,WSSVvp37和WSSV—vp39的密码子与原核生物及人的相差较大,其表达系统选择在酵母较为合适,而WSSV—vp19的密码子与原核和真核生物酵母相差较大。  相似文献   

9.
花鲈人工繁殖的研究   总被引:18,自引:0,他引:18  
1993~1995年,作者从浙江沿海三门湾及象山港采捕花鲈亲鱼,经暂养促熟处理(控制水温、盐度、溶氧水平,投喂、填喂活饵,不定期注射适量的LRH-A等),雌鱼的性腺可发育到Ⅳ_3期,对外源激素能引起正常的排卵反应。试验用的催产剂及剂量为:雌鱼每千克体重用1.5~2.5mgPG+1200~2200IUHCG+70~140μgLRH-A,或用6~10mgDOM+200~380μgLRH-A;雄鱼的注射剂量减半。在水温14.5~16℃的条件下,注射后的效应时间为36~97小时。本试验的平均受精率为89.22%,平均孵化率为75.43%。  相似文献   

10.
氨基葡萄糖盐酸盐的制备   总被引:3,自引:0,他引:3  
对以甲壳素为原料制备氨基葡萄糖盐酸盐(GAH)的工艺进行了研究,得到最佳工艺条件为:采用3.5h的水解时间;在90℃进行水解;盐酸的浓度为36%;甲壳素与盐酸的质量百分比为1:6,盐酸的回收率为60%。在此最佳工艺条件下得到的GAH经IR确认,最高产率可达36%。生物测定表明,产品GAH对大肠杆菌等菌种有一定的抗菌活性。  相似文献   

11.
饲料中添加重组鱼生长激素对罗非鱼鱼种的促生长作用研究   总被引:13,自引:2,他引:11  
采用基因工程菌EcoliDH5a(pBVGH18)生产重组鲑鱼生长激素。经初步纯化后作为添加剂投喂罗非鱼鱼种。实验结果表明,应用基因工程生产的重组鲑鱼生长激素具有生物学活性,能够通过口服经肠道被鱼体吸收,具有很强的促进鱼体生长作用。通过改进工艺,降低成本可望能作为饲料添加剂应用于水产养殖业,特别是名贵品种的养殖  相似文献   

12.
GH-transgeniccoho salmon (Oncorhynchus kitsutch) juveniles were fed diets containing 3,5,3-triiodo-L-thyronine (T3; 30 ng/g fish) or 6-n-propyl-2-thiouracil (PTU; 20 ug/g fish), to assess the effect of these drugs on the physiology, growthand survival in comparison with untreated transgenicand non-transgenic salmon. After 84 days, food intake, feed efficiency, survival, growth, hepato-somatic index (HSI), viscera-somatic index (VSI), plasma L-thyroxine (T4), T3and growth hormone (GH) levels,and cranial morphological abnormalities were determined. Growth of transgenic salmon was significantly faster than the nontransgenic salmon,and was increased by exogenous T3and reduced by PTU. Food intake of transgenic salmon was higher than that of the nontransgenic group, but was reduced by exogenous PTU administration. Food conversion efficiency of transgenic salmon was lower than that of nontransgenic salmon,and also was increased by T3 but reduced by PTU in the transgenic fish. The survival rate in all transgenic groups was significantly higher than that of nontransgenic,and transgenic T3and PTU treatment groups showed higher survivals than the transgenic-control group. The HSIand VSI of the transgenic fish were higher than the nontransgenic fish;and both parameters in the transgenic salmon were increased by PTU, but reduced by T3. The plasma T4 level in transgenic salmon was approximately 1.5-fold higher relative to the nontransgenic fish, whereas no difference was observed among the transgenic groups. Plasma T3 levels in transgenic salmon were also approximately 2-fold higher relative to the nontransgenic fish. However, the plasma T3 level in transgenic animals was increased by exogenous T3 administration, but was reduced by exogenous PTU to that observed in nontransgenic salmon. The plasma GH level of transgenic fish was higher than that of the nontransgenic salmon,and the level was increased by the exogenous T3, whereas exogenous PTU did not reduce significantly GH levels in transgenic salmon. Transgenic fish also displayed cranium, jawand opercular abnormalities typical of the effects of this gene construct incoho salmon, indicating that some imbalance in growth processes has been induced. However, these abnormalities (especially cranial disruptions) were diminished by administration of exogenous PTU. In conclusion, exogenous T3and PTU treatments can induce hyperthyroidismand hypothyroidism, respectively,and have inverse effects on growthand skeletal abnormalities of transgenic salmon constitutively expressing GH.  相似文献   

13.
A feeding trial was conducted to evaluate the results of replacing soybean meal (SBM) with other blend plant protein (BP) (rapeseed meal, cottonseed meal and peanut meal) sources on growth, fish body composition, biochemical parameters, non‐specific immune index and gene expression of growth hormone/insulin‐like growth factor‐1 in Yellow River carp Cyprinus carpio. The results showed that the 600 g/kg replacement with BP in diet did not affect the growth performance of Yellow River carp, but the 800 g/kg SBM replacement diet could depress the growth of fish (p < .05). The 800 g/kg SBM replacement diet significantly impacted the superoxide dismutase, malondialdehyde contents, lysozyme, alanine aminotransferase and aspartate aminotransferase activities (p < .05). Fish fed 800 g/kg SBM replacement diet showed lowest insulin‐like growth factor 1 and highest growth hormone level than that of other diets (p < .05). Both gene expression levels of GH and IGF‐I in hepatic showed significant difference among dietary treatments (p < .05), and the lowest GH and IGF‐I mRNA level in liver were found in fish fed 800 g/kg SBM replacement diet (p < .05). In conclusion, no more than 600 g/kg SBM could be replaced by BP in diet of Yellow River carp without adverse effects on the growth performance. However, 800 g/kg SBM replacement with BP in diet inhibited the growth performance, affected some blood parameters and immune response index, and down‐regulated GH and IGF‐1 gene expression of liver in Yellow River carp. Further, GH level in serum was negatively correlated with GH mRNA level in liver; meanwhile, serum concentrations of IGF‐I were positively correlated with hepatic IGF‐I mRNA expressions.  相似文献   

14.
Molecular cloning and expression of salmon pituitary hormones   总被引:2,自引:0,他引:2  
A cDNA library was prepared from chinook salmon pituitaries. Growth hormone (GH), prolactin (PRL) and the β subunit of gonadotropin (GTH) genes were screened using synthetic oligonucleotides as probes. Full size cDNA clones coding for these polypeptide hormones were isolated and characterized. The cDNA sequences for PRL and βGTH have been reported earlier from our laboratories. The cDNA clone for GH contains 1148 bp and codes for a preGH of 210 amino acids. The chinook salmon GH, reported in the present investigation, differs from chum salmon GH in only 1 amino acid, and from coho salmon GH in 5 amino acids. Plasmids containing modified nucleotide sequences coding for GH, PRL and βGTH were constructed individually into an expression vector using the heat-inducible λ pL promotor. Mature PRL, GH and unglycosylated βGTH were expressed in the bacteria at elevated temperature.  相似文献   

15.
In salmonids, growth hormone (GH) stimulates growth, appetite and the ability to compete for food. This study tested the hypothesis that increased GH levels in GH-transgenic coho salmon Oncorhynchus kisutch (Walbaum) increase competitive ability through higher feeding motivation. The transgenic strain of salmon used contained a gene construct consisting of the sockeye metallothionein-B promoter fused to the type 1 growth gene coding region. The transgenic animals (mean size = 250 g) were F1 individuals. In six consecutive feeding trials, the intake of contested food pellets by size-matched pairs consisting of one control (1 year older non-transgenic coho salmon) and one GH-transgenic coho salmon was compared. Pellets were provided sequentially until neither fish took three consecutive pellets; the identity of the fish taking each pellet was noted. Calculated on the three first pellets offered at each feeding trial, the transgenic coho salmon consumed 2.5 times more contested pellets than the controls, supporting the hypothesis that GH transgenesis increases the ability to compete for food. Overall, the transgenic fish consumed 2.9 times more pellets that the non-transgenic controls, indicating a high feeding motivation of the transgenic fish throughout the feeding trials. It appears that GH transgenesis and GH treatments can induce similar changes in the feeding behaviour of salmonids. Depending on how transgenic and wild individuals differ in other fitness-related characters, escaped GH transgenic fish may compete successfully with native fish in the wild.  相似文献   

16.
王雪惠 《水产学报》2006,30(3):297-304
采用RACE-PCR技术结合SMART cDNA合成技术,从银鲫中克隆到Ran的全长cDNA并对其编码区全长进行了原核表达、相应抗体制备及其时空表达特征分析。RT-PCR结果表明,Ran基因除在脑组织的转录水平较低外,其它组织中的转录水平几乎相同;Ran基因在不同发育阶段的胚胎中都有mRNA转录。但其mRNA的量在原肠期以后呈下降趋势。Western blot结果表明,Ran在卵巢和精巢中均高水平表达,在心、脑、肝、脾、肾中有较低水平表达,在肌肉中则不表达。同时检测到Ran在不同胚胎发育阶段均有较强表达。这表明,Ran基因在银鲫中可能起着多种作用,尤其在生殖细胞的发生中具有重要作用。最后,采用免疫耗竭对该基因编码的蛋白在精核解凝中的作用进行了初步研究,结果表明,Ran蛋白可能与精核解凝相关。  相似文献   

17.
孙颖 《水产学报》2006,30(6):740-746
家蚕多角体病毒表达系统在家蚕体内表达基因重组的草鱼生长激素(r-gGH),具有与天然草鱼生长激素(GH)相似的免疫原性和生物活性。为了节省基因工程研究中的下游工作 (基因产物的分离和提纯),本研究将含有r-gGH的家蚕直接作为饵料源,冻干并磨碎后拌在饵料中投喂草鱼鱼种,通过养殖实验及生化测定分析对比r-gGH促进草鱼鱼种生长的剂量依存关系,筛选活性强的处理剂量和处理时间,期望为鱼类养殖生产提供一种较为经济、来源容易、方法简易而又切实可行的促进鱼种生长并且可以大规模应用的方式。实验结果表明,投喂含有r-gGH的家蚕,有相当一部分被鱼体消化道吸收,进入血液循环。投喂2 h和6 h 后,草鱼鱼种的血清GH水平均显著高于对照组(投喂基本饲料)和投喂正常家蚕组;每天投喂和隔2天投喂,均使草鱼鱼种的血清GH水平显著升高,并对草鱼鱼种生长都有明显的促进作用。短期(3 d)和长期(42 d)投喂含有r-gGH 的家蚕,无论是低剂量(10 mg·g-1饲料)还是高剂量(20 mg·g-1饲料),均极其显著地提高草鱼鱼种的血清GH水平;长期(42 d)投喂亦对草鱼鱼种的生长有显著的促进作用,鱼体的相对体重增长率、相对体长增长率、食物转化率和肥满度显著增加。  相似文献   

18.
Since somatostatin (SRIF) inhibits the release of growth hormone (GH), its immunoneutralization may provide an alternative to GH therapy as a means of enhancing somatic growth in fish. The present study examined the feasibility of accelerating growth in juvenile chinook salmon by means of antiSRIF administration. Yearling salmon of Nicola River stock (BC, Canada) were injected intraperitoneally every 5 days, for a total of 40 days, with either SRIF (1 μg g-1 body wt.), antiSRIF (SOMA-10, 1 μg g−1), recombinant bovine GH (rbGH, 2.5 μg g−1), recombinant porcine GH (rpGH, 2.5 μg g−1) or saline (controls). No significant differences were observed in length, weight or final condition factor (k) between the SRIF-treated and control fish over the experimental period. However, the fish treated with the antiSRIF were significantly (p ≤ 0.05) longer and heavier than the control salmon after 25 and 30 days respectively. Furthermore, antiSRIF treatment caused a lowering in k when compared to the control salmon. Fish injected with rbGH or rpGH were significantly longer and heavier than all other groups (p ≤ 0.05), after only 5 days. GH treated groups also returned higher k when compared against all other treatments (p ≤ 0.05). No differences were observed in growth between the two rGH treatments over the experimental period.  相似文献   

19.
Two marine algal products MAP3 and MAP8 were examined for their suitability as fishmeal protein substitutes in feeds of three prominent farmed species, through short‐term feeding studies. Algal meals were tested at 5 and 10% protein replacement levels for Atlantic salmon and at 25 and 40% for common carp and whiteleg shrimp. At the end of the 12‐week period, the growth and feed performance of the two fish species did not reveal any significant difference between those fish offered the algae‐based feed and those offered the control feed. The whole body proximate compositions of Atlantic salmon fed the control and algae‐based feeds were not significantly different. In common carp, the lipid content in the fish fed higher level of MAP3 was significantly lower than that of the fish fed the control feed. In whiteleg shrimp, at the end of the 9‐week feeding period, growth performance and feed utilization did not differ between the treatment groups. Protein content in the shrimp fed the higher level of MAP8 was significantly lower than that of shrimp on the control feed. The three species could accept the algal meals in their feeds at the tested levels, though there were some noticeable effects on body composition at higher inclusion levels.  相似文献   

20.
采用RACE-PCR方法,从鲢(Hypophthalmichthys molitrix)头肾SMARTcDNA中获得了679 bp IL-8 cDNA序列。结果显示:该序列包含169 bp的5′非编码区,213 bp 3′端非编码区和297 bp的开放阅读框;鲢IL-8的开放阅读框编码98个氨基酸残基,其中包含构成两对二硫键的4个保守半胱氨酸。RT-PCR结果显示鲢:IL-8 mRNA主要在胰、肝脏和头肾中表达。将鲢IL-8不含信号肽的成熟多肽克隆到原核融合表达载体pQE30上,转入大肠杆菌M15内进行表达,经SDS-PAGE电泳后显示,重组融合蛋白在分子量约11kD处有一明显表达带,与预期计算的分子量大小相一致。  相似文献   

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