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1.
取本室构建的鲤生长激素毕赤酵母工程菌GS115 (pPICZαA -GH) ,甲醇诱导 ,SDS -PAGE检测 ,证明重组鲤生长激素 (RecombinantCarpGrowthHormone ,rcGH)在酵母中得到分泌表达。重组酵母菌生长曲线显示 ,细胞从培养第 8小时进入对数生长期 ,至 32hOD660 升至峰值 11.5 ,此后OD660 缓慢下降。在以 10 0 %甲醛按 0 .5 %添加量诱导至 72h、pH为 6 .0时 ,rcGH表达量最高。凝胶扫描分析 ,分泌表达的rcGH占上清总蛋白的 36 .4 1% ,表达量为 30 0～ 4 0 0mg/L发酵液。经离子交换层析法纯化表达产物 ,纯度可达 95 %。纯化的rcGH注射奥尼罗非鱼 ,结果表明具有明显的促生长作用 相似文献
2.
根据红鳍东方鲀生长激素基因全序列,设计13对引物,结合单链构象多态性(SSCP)技术和克隆测序技术对野生红鳍东方鲀、养殖红鳍东方鲀和养殖暗纹东方鲀群体进行了遗传多样性分析.结果表明:东方鲀整个生长激素基因的6个外显子中没有多态性,在其内含子2、内含子5以及3′侧翼区发现有多处序列变异位点,包括碱基的缺失、插入和一些单核苷酸突变.这些遗传多态性的存在为从分子水平上对东方鲀的鉴定提供了一定的依据.群体遗传结构分析结果表明,红鳍东方鲀养殖群体的一些等位基因丧失,造成其遗传多样性显著下降,应及早加强对其种质的保护. 相似文献
3.
采用聚合酶链反应（ＰＣＲ）技术对鲤和鲑生长激素ｃＤＮＡ的５‘端和３’端进行定向改造。将改造后的５种基因分别克隆到大肠杆菌表达质粒ｐＢＶ２２０进行原核表达,以研究鱼生长激素基因在大肠杆菌中的表达水平。实验结果表明：（１）核糖核蛋白体结合位点（ＳＤ）与起始密码子ＡＵＧ之间的距离对鱼生长激素的表达水平有影响;（２）鲑鱼生长激素基因的终止密码子ＵＡＧ可能不会有效终止该基因在大肠杆菌中翻译的进行而造成部分通 相似文献
4.
Growth hormone (GH) and reproduction: a review 总被引：5，自引：0，他引：5
Florence Le Gac Odile Blaise Alex Fostier Pierre-Yves Le Bail Maurice Loir Brigitte Mourot Claudine Weil 《Fish physiology and biochemistry》1993,11(1-6):219-232
Interaction between growth and reproduction occurs in many vertebrates and is particularly obvious at certain stages of the life cycle in fish. Endocrine interactions between the gonadotropic axis and the somatotropic axis are described, the potential role of GH being emphasised. A comparative analysis of these phenomena in mammals, amphibians and fish, suggests a specific role of GH in the physiology of puberty, gametogenesis and fertility. It also shows the original contribution made by studies on the fish model in this field of investigations. 相似文献
5.
Growth hormone (GH) has recently been identified as co-gonadotropin regulating fish reproduction, hitherto, no effort has
been made to see its effect on oocyte maturation in fishes, though some reports demonstrate the role of insulin like growth
factor-I (IGF-I) in oocyte maturation in teleosts. Hence, effect of GH on oocyte maturation in post-vitellogenic H. fossilis has been worked out in the present study. Post-vitellogenic follicles in the ovarian tissue were challenged in vitro with H. fossilis pituitary homogenate (fPH), Clarias batrachus GH and GtH, barramundi IGF-I (IGF-I), 17α, 20β-dihydroxy-4-pregnen-3-one (DHP) and testosterone alone, or in combination with IGF-I for 18 h at 26±1°C. Incubation of ovarian
tissue with GH in the presence of actinomycin d or cycloheximide or barramundi IGF-I antiserum was also made separately. In general, oocyte maturation was induced by fPH,
barramundi IGF-I, GtH, GH and DHP, which was augmented further by addition of barramundi IGF-I. Testosterone had no effect
on GVBD. Actinomycin d, cycloheximide and anti barramundi IGF-I abolished the GH induced oocyte maturation. Present study suggests for the first
time that GH has a role in egg maturation in fish. 相似文献
6.
利用长片段PCR及T-A克隆技术,从兰州鲇(Silurus lanzhouensis)基因组DNA中首次克隆兰州鲇GH基因全序列并提交至Genbank获得登录号KM215221,同时,使用DNAstar等生物信息学软件进行序列分析研究。兰州鲇GH基因全长2 067 bp,由5个外显子和4个内含子组成,完整编码序列(Complete coding sequence,CDS)为603 bp。5个外显子大小分别为10、140、117、132和204 bp;4个内含子大小分别为229、103、565和103 bp;内含子与外显子的连接区序列遵循基因组成规则。编码区编码1条由200个氨基酸残基组成的蛋白质多肽。5个科8种鲶形目鱼类生长激素基因编码区序列同源性比较表明,它们之间GH基因编码区序列有较高的同源性,平均达到了92.4%,其中,兰州鲇和南方大口鲇之间的同源性最高,为99.5%,兰州鲇和革胡子鲇之间的同源性最低,为90.0%,并构建了系统发育进化树。同时也表明,GH基因编码区序列具有较高的保守性,系统发育研究中具有较高的参考价值。 相似文献
7.
Pierre-Yves Le Bail Geneviève Boulard Bruno Barenton Michel Zygmunt 《Fish physiology and biochemistry》1989,7(1-6):243-251
A method for the purification of chinook Salmon (Oncorhynchus tshawytscha) GH, which retains its biological activity, is described. The biological activity was investigated with an established radioreceptor
assay using liver membranes from pregnant rabbits and bovine GH as standard and labelled hormone. The enrichment of the preparation
was checked with electrophoresis (SDS-PAGE). Extraction and further steps were carried out using low molarity alkaline buffer
(pH 8–10, M = 100 mM). Three chromatography steps were performed (Concanavalin-A sepharose, Bio-gel P60, DEAE). Ion exchange
chromatography was performed under isocratic conditions (using a 50 cm column). Two isoforms (sGH1 and sGH2) were isolated.
The purification yield is 0.7% compared to lyophilized pituitaries. The molecule is homogeneous in SDS-PAGE. Contamination
by prolactin, gonadotrophin and corticotrophin is negligible (< 0.5%). It could be demonstrated that the biological activity
of the preparation is maintained since this preparation stimulates the growth of juvenile trout (Salmo gairdneri) and binds specifically (35%) to trout liver membranes. 相似文献
8.
Ghrelin was recently demonstrated as an endogenous ligand of the growth hormone (GH) secretagogue receptor (GHS-R), which could promote the release of GH in mammal significantly. The present study conducted to determine whether ghrelin stimulate the release and synthesis of GH in orange-spotted grouper (Epinephelus coioides). Rat ghrelin was incubated with the pituitary fragments of grouper in static culture system. The culture medium was collected at 1, 6, 12, 18 and 24 h after incubation to detect the contents of GH by homologous radioimmunoassay. The level of GH mRNA in the pituitary fragments was measured by a sensitive chemiluminescent ribonuclease protection assay. The results showed that rat ghrelin not only stimulated the release of GH but also augmented the GH mRNA level in grouper. It suggested that the ghrelin-like peptide and the GHS-R involved in the regulation of GH synthesis and release in grouper. The present study would provide a better understanding of the regulatory mechanism of GH release in marine fish. 相似文献
9.
生长激素（ GH）基因能够影响动物的生长,是一个与生长性状有关的基因。以315尾甘肃金鳟（ Oncorhyn-chus mykiss）为研究材料,分析了GH基因外显子区域多态性,并对基因型与生长性状进行了关联性分析。 PCR-SSCP分析结果显示B等位基因在外显子2区域第903 bp处发生G/C突变,存在3种基因型AA、 AB和BB。 D等位基因在外显子5区域第3049 bp处发生T/G的突变,在3094 bp处发生C/A的突变,其中第3049 bp处发生的T/G的突变导致第157位氨基酸由组氨酸变为谷氨酰胺,为有义突变,共检测到3种基因型CC、 CD和DD个体。最小二乘法分析表明, BB基因型的全长值和体长值都显著高于AA和AB基因型（ P＜0.05）。 T3049 G和C3094 A突变点对全长、体长和体重均有显著性影响（ P＜0.05）,其中CC基因型的全长值、体长值和体重值都高于DD和CD基因型。在标记辅助育种中,这些分子标记可以为甘肃金鳟遗传改良提供基础的数据和参考。 相似文献
10.
11.
本研究旨在克隆梭鲈(Sander lucioperca)生长激素(growth hormone, GH)基因, 探究其序列特征及其表达量与梭鲈生长的相关性, 以期为梭鲈生长的分子机理研究鉴定基础。研究采用 RACE 技术从梭鲈垂体中克隆了 GH 基因, 其 cDNA 全长为 926 bp, 包含 74 bp 的 5′非翻译区、237 bp 的 3′非翻译区和编码 204 个氨基酸的开放阅读框。 氨基酸多重比对发现, 梭鲈 GH 与黄金鲈(Perca flavescens)、河鲈(Perca fluviatilis)的氨基酸序列相似性分别为 97.04%和 94.05%, GH 基因在鱼类和哺乳动物中都存在四个保守的半胱氨酸残基位点。应用实时荧光定量 PCR 技术分析了 GH 基因在梭鲈各组织和体重极端差异个体间的表达特征, 结果显示, GH 基因在梭鲈各组织中均有表达, 垂体中相对表达量最高, 脑次之, 与其他组织相比差异极显著(P＜0.01)。GH 基因表达量与梭鲈体重呈正相关, 4 个组织中 GH 基因表达量均为体重极大组高于极小组, 其中垂体和脑中差异极显著(P＜0.01), 肌肉中差异显著 (P＜0.05)。因此, GH 基因可以作为梭鲈生长候选基因用于分子选择。本研究结果可为进一步解析 GH 基因调控梭鲈生长发育的分子机制研究提供参考。 相似文献
12.
使用半定量RT-PCR方法,研究了鲮（Cirrhinus molitorella）各组织中IGF-ⅠmRNA组织表达,并分析重组鲮GH处理后鲮IGF-Ⅰ表达变化情况。结果表明,鲮IGF-ⅠmRNA在肝组织中表达最高,其次是肾和脑,另外在肠、鳃、脾、性腺和心脏组织中也有表达,而在肌肉、皮肤中没有检测到鲮IGF-ⅠmRNA的表达;按120μg·g^-1鱼体重的剂量经腹腔注射重组鲮GH,观察12h后鲮IGF-Ⅰ表达变化情况,发现经重组鲮GH处理后,鲮肝组织IGF-ⅠmRNA表达水平显著升高,脑组织IGF-ⅠmRNA表达水平也稍有升高,而在肌肉中仍未检测到IGF-ⅠmRNA的表达;重组鲮GH处理前后,对照组中鲮血清中IGF-Ⅰ的含量为145.59±21.84ng·mL^-1,试验组中鲮血清中IGF-Ⅰ的含量为247.71±2.83ng·mL^-1,经t检验,P=0.043〈0.05,表明重组鲮GH处理前后鲮血清中IGF-Ⅰ的含量存在显著性差异。 相似文献
13.
Effect of diet and ration on the relationship between plasma GH and IGF-1 concentrations in Arctic charr, Salvelinus alpinus (L.) 总被引：1，自引：0，他引：1
Colin Cameron Richard Moccia Paula A Azevedo & John F Leatherland 《Aquaculture Research》2007,38(8):877-886
The purpose of the study was to investigate whether dietary ration or diet composition influence the relationship between plasma growth hormone (GH) and insulin‐like growth factor‐1 (IGF‐1) in Arctic charr (Salvelinus alpinus L.). The pattern of changes in plasma GH and IGF‐1 concentrations was examined in fish fed at different ration levels (0%, 0.35% and 0.70% BW day−1) for 5 weeks, and in fish fed diets containing different lipid:crude protein (LCP) ratios. Ration level significantly affected plasma GH and IGF‐1 concentrations; at 5 weeks the levels of both hormones in the food‐deprived group were significantly lower than in fish fed the 0.70% BW day−1 ration. Also, plasma IGF‐1 levels in fish of each ration treatment group were significantly correlated with individual final body weight; no such correlation was found for GH. To examine the effects of dietary LCP ratios, fish were fed for up to 18 weeks, with one of four formulated diets that had LCP ratios (dry matter basis) of 0.35 (Diet 1), 0.43 (Diet 2), 0.51 (Diet 3) or 0.59 (Diet 4), or a commercial diet (Diet 5) which had an LCP ratio of 0.38. Statistical differences in plasma GH and IGF‐1 concentrations were found only after 18 weeks. Growth hormone was significantly lower in fish fed Diets 1 and 2 compared with Diets 3 and 5, and IGF‐1 was significantly lower in fish fed Diet 1 compared with Diets 2 and 5. Significant correlations between plasma GH and IGF‐1 concentrations were found only for fish fed Diets 1 and 5, suggesting that the influence of diet composition on the relationship between GH and IGF‐1 varies with the dietary LCP ratio in this species. The decline in plasma IGF‐1 concentrations during food deprivation is similar to that described in other species; however, the unexpected decrease in plasma GH during food deprivation in this study may represent a species‐specific response. 相似文献
14.
European eels reaching the silver stage stop feeding in freshwater and during their spawning migration to the Sargasso sea (6000 km at least in the Ocean). The total duration of this exceptional fast is not well known. Few data are available on the general condition and the endocrine responses to starvation of migrating eels. In this study male (silver) and female (yellow and silver) eels were kept in freshwater without food for various lengths of time. Animals were killed after 7 months to 3 or 4 years. After a gradual decrease, the final body weight was reduced by 84% in males (52 months) and 67–69% in females (up to 4 years). The condition factor (K) followed a parallel curve. In the pituitary gland, GH cells were hypertrophied and highly stimulated. Their cross-sectional area was negatively correlated to K. Large GH cells remained well immunostained with an anti-eel GH serum after 7 to 12 months of starvation. In the leanest eels, the immunostaining was often reduced and many GH cells appeared degranulated, suggesting a low hormonal storage. In contrast, SL cells were reduced in size and number in the anterior half of the neurointermediate lobe (NIL), but showed a more heterogeneous picture in the caudal portion. GH and SL cell activities seemed to be negatively correlated in starved fish and controlled by different mechanisms. GH appears to play a major role in the long-term survival of fasted eels; SL does not seem to be involved. 相似文献
15.
在室内可控条件下,对尼罗罗非鱼[初始体质量(62.50±3.44)g]进行饥饿28d和随后再投喂21d的处理,于饥饿第0、7、14、21、28天和再投喂第14、21天进行采样分析,研究饥饿和再投喂期间尼罗罗非鱼生长、血清生化指标和肝胰脏生长激素(GH)、类胰岛素生长因子-Ⅰ(IGF-Ⅰ)和胰岛素(IN)mRNA表达丰度的变化。结果显示,与饥饿第0天相比,饥饿超过7d鱼体体质量显著降低(P<0.05),再投喂21d显著增加(P<0.05);肝体比随饥饿时间延长显著降低(P<0.05),恢复投喂后较饥饿时升高,但显著低于饥饿前水平(P<0.05)。在血清指标上,甘油三酯、血糖、碱性磷酸酶、谷草转氨酶和谷丙转氨酶均随饥饿时间延长而逐渐降低,恢复投喂后均有不同程度提高,但转氨酶活性显著低于饥饿前水平(P<0.05);饥饿和再投喂对血清总胆固醇、高密度脂蛋白胆固醇和低密度脂蛋白胆固醇无显著影响(P>0.05)。在激素方面,与饥饿第0天相比,饥饿使血清GH含量及其肝胰脏mRNA表达丰度显著升高,血清IGF-Ⅰ及其肝胰脏mRNA表达丰度降低,恢复投喂后两者均显著升高(P<0.05);INmRNA表达丰度在饥饿7~21d显著升高(P<0.05),饥饿第28天时无显著差异(P>0.05),再投喂后显著降低(P<0.05)。 相似文献
16.
We have studied the seasonal relationship between growth and circulating growth hormone (GH), hepatic GH-binding and plasma
insulin-like growth factor-I immunoreactivity in gilthead sea bream,Sparus aurata. The seasonal increase in plasma GH levels preceded by several weeks the summer increase in growth rates. In contrast, a
marked increase in hepatic GH-binding with a high degree of endogenous GH occupancy was found during the period of maximum
growth which suggests an enhanced sensitivity of liver to GH action. Thus, circulating levels of immunoreactive IGF-I, probably
derived from the liver in response to GH action, were positively correlated with growth throughout the experimental period
although a consistent relationship between growth and circulating GH was not found. In spite of this, we consider that, in
gilthead sea bream, as in several other teleosts, the availability of endogenous GH can limit growth. Thus, under environmental
conditions of suboptimal growth, a single intraperitoneal injection of recombinant rainbow trout GH (rtGH) induced over the
dose range tested (0.75, 1.5, 3 μg g BW−1) an increase in plasma IGF-I-like immunoreactivity comparable to that seen during the period of maximum growth. 相似文献
17.
Kouakou Yao Jean-Charles Gabillard Pierre-Yves Le Bail 《Fish physiology and biochemistry》2006,32(2):121-130
In fish, regulation of the growth hormone (GH) receptor has been mainly analyzed by binding studies, with some discrepancies in the results. The present work aims at determining whether circulating GH levels influence the measurement of hepatic GH-binding capacities. To do this, the effectiveness of the dissociation of the GH/GH receptor (GHR) complex by an MgCl2 treatment was assessed in rainbow trout, and data on GH-binding capacities under various physiological conditions were studied using different means of expression. Our results reveal that MgCl2 treatment dissociated the liver GH/GHR complex formed under in vitro conditions (85 ± 23 vs. 361 ± 16 fmol/g of liver; p<0.001) but not in vivo, showing such treatment in trout is not applicable. A comparison of fasted (3 weeks) and fed fish revealed that GH-binding capacities, expressed as femtomoles per milligram protein or femtomoles per gram of liver, were similar in both fed and fasted fish. However, when changes in liver and body size were taken into account, the total GH-binding capacities were lower in the fasted fish (0.026 ± 0.006 vs. 0.062 ± 0.009 fmol/cm3 liver; p<0.05). One day after hypophysectomy or GH injection, changes in the plasma GH levels increased or decreased GH-binding capacities, respectively. Five days later, GH-binding capacities increased in GH-injected fish (527 ± 38 vs. 399 ± 38 fmol/g liver; p<0.01). Our interpretation is that acute treatment modified GH-binding capacities through receptor occupancy and that GH stimulated the synthesis of its own receptor. On the other hand, long-term treatment through successive injections of GH lowered the total binding capacities (approx. 40%), which could result from receptor occupancy. We conclude that circulating GH levels strongly influence the measurement of GH-binding capacities in the liver, thereby limiting interpretation of the binding data and preventing accurate conclusions to be drawn on GHR regulation. 相似文献
18.
褐牙鲆（Paralichthysolivaceus）幼鱼分别在温度8．5℃（T8.5）、13．0℃（T13.0）、17．5℃（T17.5）、22．0℃（T22.0）、26．5℃（T26.5）下养殖10d后,调整至生长最快的温度22．0℃养殖30d,研究了生长激素（GH）、类胰岛素生长因子I（IGF-I）、RNA／DNA比值和糖原质量分数的变化。除在胁迫阶段T17.5处理血浆IGF-I质量浓度显著低于T22.0处理外,不同处理的血浆GH和IGF-I质量浓度在胁迫和恢复阶段均无显著差异。不同处理的肝脏RNA／DNA比值仅在20-40d阶段有显著差异,不同处理的肌肉RNA／DNA比值在所有时间均无显著差异。胁迫结束时T8.5、T13.0、T26．5处理的肝脏糖原质量分数显著低于T17.5,处理,试验结束时T22.0处理肝糖原质量分数略低于其余处理。胁迫结束时T25.5处理肌肉糖原质量分数最高,而在恢复的第一个10d时T13.0处理肌肉糖原质量分数最高。不同温度下养殖10d导致的生长减缓能够在恢复至22．0℃后的30d内获得完全补偿生长,但此试验中GH、IGF-I和RNA／DNA比值与生长率不存在明显的相关性。 相似文献
19.
20.
投喂频率对池塘工程化循环水养殖大口黑鲈生长、生理及肝脏GH、IGF-I基因表达丰度的影响 总被引：1，自引：1，他引：0
为研究池塘工程化循环水养殖模式下投喂频率对大口黑鲈(Micropterussalmoides)生长、生理及肝脏生长激素基因(GH)、类胰岛素生长因子-I基因(IGF-I)相对表达丰度的影响,选用初始体重为(5.0±0.4) g的大口黑鲈为研究对象,设置3个投喂频率组(2次/d、3次/d、4次/d),每组3个重复,开展为期120 d的养殖试验。结果表明,第30天和第60天时,大口黑鲈末体重、增重率和特定生长率均未受到投喂频率的显著影响(P0.05),而在第90天时, 2次/d组试验鱼的生长显著高于4次/d组(P0.05),与3次/d组相比无显著性差异(P0.05);第120天时, 2次/d组试验鱼的生长显著高于3次/d组和4次/d组(P0.05);投喂频率对血清谷氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST)、碱性磷酸酶(ALP)活性、溶菌酶、皮质醇和甘油三酯(TG)含量、肝脏总抗氧化能力(T-AOC)、谷胱甘肽过氧化物酶(GSH-Px)活性及丙二醛(MDA)含量均无显著性影响(P0.05)。随着投喂频率的增加,肝脏过氧化氢酶(CAT)活性及血清总蛋白(TP)、血糖(Glu)含量呈降低的趋势且血清总胆固醇(TC)呈上升的趋势;第30天时投喂频率对肝脏总超氧化物歧化酶(T-SOD)活性无显著性影响,而第60天、第90天和第120天时,4次/d组T-SOD活性显著低于2次/d组(P0.05)。第30天、第60天和第120天时, 2次/d组肝脏IGF-I基因的相对表达量最高(P0.05),第90天时各组间肝脏IGF-I相对表达量无明显差异(P0.05);第30天和第60天时,肝脏GH基因相对表达量未受到投喂频率的影响(P0.05),而第90天和第120天时4次/d组肝脏GH基因相对表达量显著低于2次/d组(P0.05)。因此,基于大口黑鲈的生长性能、生理效应及肝脏GH、IGF-I基因表达丰度的综合考虑,池塘工程化循环水养殖模式下初始体重为(5.0±0.4)g的大口黑鲈适宜投喂频率为2次/d。 相似文献