共查询到18条相似文献，搜索用时 218 毫秒
1.
《水产学报》2021,(6)
为探究大黄鱼在高温胁迫条件下的蛋白表达情况,实验应用串联质谱标签tandem mass tag (TMT)标记结合液相色谱-串联质谱(LC-MS/MS)技术对大黄鱼耐高温组和不耐高温组的肝脏组织进行蛋白组学分析。共鉴定到3 369个蛋白,其中有687个差异表达蛋白,包括281个上调蛋白和406个下调蛋白。用平行反应监测(parallel reaction monitoring, PRM)对随机挑选的13个蛋白进行验证,其结果与TMT标记的蛋白组学分析结果一致。随后,通过生物信息学分析发现,高温胁迫对大黄鱼的蛋白质折叠、能量代谢有显著影响,其中热休克蛋白70 (heat shock proteins 70, HSP70)、钙网蛋白(calreticulin,CRT)和葡萄糖调节蛋白(glucose-regulated protein, GRP)参与调节蛋白质的正确折叠,丙酮酸激酶(pyruvate kinase, PK)参与高温条件下的能量供给。由此推测HSP70、CRT、GRP和PK在大黄鱼应对高温胁迫时发挥着重要的作用。 相似文献
2.
葡萄糖调节蛋白78(glucose regulated protein 78 ku,GRP78)是热休克蛋白70家族成员之一,在调节蛋白质折叠和维持内质网稳态过程中起着分子伴 侣作用。采用RT-PCR、RACE等技术,首次从拟穴青蟹获得了GRP78的cDNA全长序列。该序列全长2 284 bp,开放阅读框(ORF)为1 962 bp,编码653个氨基酸残基。 同源分析显示,该基因编码的蛋白含有HSP70家族的签名序列,C末端为内质网蛋白滞留信号KDEL,与其他物种具有很高相似性。实时荧光定量PCR结果表明,GRP78 基因在拟穴青蟹多个组织中均有表达。第一期仔蟹在不同的温度和盐度下暴露12 h后,GRP78基因表达量随环境温度升高而增加;在高盐(30)条件下GRP78表达量 较高,进而推测拟穴青蟹GRP78参与蛋白质折叠和环境胁迫的应答。 相似文献
3.
本研究以海带(Saccharina japonica)‘海天1号’为材料,通过RACE-PCR方法获得海带hsp70(Sjhsp70)基因全长cDNA序列。序列分析表明,全长cDNA长度为3778 bp,含有1个2892 bp的开放阅读框,5'和3'非编码区的长度分别为101、785 bp。蛋白质氨基酸含量和理化性质分析显示,该蛋白为稳定蛋白。应用BLASTP程序对不同物种HSP70蛋白比较分析表明,海带与长囊水云(Ectocarpus siliculosus)的HSP70蛋白具有较高的同源性,用HSP70蛋白构建的进化树显示,海带与长囊水云聚为一类。二级结构预测该蛋白由963个氨基酸组成,包含35个α螺旋和25个β折叠。以酿酒酵母(Saccharomyces cerevisiae)HSP110蛋白三维结构3C7N为模板,在I-TASSER软件上预测海带HSP70蛋白的三维结构,预测的蛋白结构与3C7N的A链在折叠和二级结构方面非常类似。对Sjhsp70基因在温度胁迫下表达变化分析表明,高温对Sjhsp70的表达量具有显著影响,Sjhsp70在25℃、24 h胁迫条件时,表达量达到最大值。本研究为阐明海带HSP70蛋白特性及其应对环境胁迫的机理提供理论依据。 相似文献
4.
大黄鱼高温适应的转录组学分析 总被引：3，自引：3，他引：0
为了探究大黄鱼高温胁迫条件下基因表达水平的变化,实验利用Illumina Hiseq 2500的125 pair-ended测序模式分别对大黄鱼高温处理组和常温对照组进行了转录组测序。对照组(3个生物学重复)和高温处理组(3个生物学重复)分别获得15.28和13.92 Gb测序数据,GC含量平均值约为51%。过滤后的高质量测序reads使用Bowtie2软件比对到大黄鱼参考转录组序列上估计基因表达量,进而进行基因表达差异统计学检验。以常温对照组为参比,高温处理组中阈值设为|log2(FC)|2和FDR0.05,共检测到1 259条显著差异表达基因。其中,821条基因为高表达,438条基因为低表达。随机选取12条差异表达基因,运用实时荧光定量PCR(qRT-PCR)进行了验证,结果证实转录组分析可靠。进一步将所有差异表达的基因进行GO功能注释和KEGG通路富集分析,结果发现大量差异表达基因的功能与氧化还原反应、蛋白质折叠和去折叠、糖和脂代谢以及某些疾病的发生相关。该研究结果为下一步深入研究大黄鱼高温适应的调控机制提供了重要的参考材料。 相似文献
5.
6.
为从蛋白角度探索红罗非鱼(Oreochromis mossambicus♀×O.niloticus♂)在盐胁迫环境下的调节适应机制,本研究采用蛋白抗体芯片结合串联质谱技术初步筛选了盐胁迫下红罗非鱼鳃组织的差异表达蛋白,并通过免疫组化和免疫印迹技术对候选差异蛋白进行了验证。结果显示,共筛选获得181个表达量有显著差异的蛋白质(变化倍数≥1.5),包含上调蛋白142个,下调蛋白39个。挑选5个蛋白进行质谱鉴定得到3个蛋白质:中间丝蛋白(intermediate filament protein,IF)、转运蛋白63(translocation protein 63,SEC63)、蛋白质二硫键异构酶A3(protein disulfide-isomerase A3,PDIA3)。免疫组化结果显示,在淡水、盐度组中,IF和PDIA3在鳃小片基部均有阳性反应,且随盐度升高,阳性反应呈先降低后增强的趋势,SEC63无阳性反应。Western blot结果显示,在淡水和盐度组中,IF和PDIA3两种蛋白均有表达,并且随盐度升高呈现先降低、再升高的变化趋势,SEC63蛋白无明显目的条带;在30盐度组中,胁迫早期,IF蛋白表达量降低,48 h达到最低值,72 h有明显回升,PDIA3蛋白表达量在胁迫后96 h显著升高(P0.05)。根据研究结果推测,IF和PDIA3是在盐胁迫环境下红罗非鱼鳃组织的响应蛋白,它们分别在细胞骨架、内质网功能维持中发挥重要调节作用。 相似文献
7.
为探讨铜驯化对高温胁迫下大黄鱼(Larimichthys crocea)氧化损伤和基因表达水平的影响,将体质量为(48.92±3.62) g的大黄鱼暴露在铜离子浓度为0和10μg·L-1的水体中14 d,再暴露在温度为32℃的水体中24 h。结果显示,高温胁迫显著增加了大黄鱼肝脏中活性氧(ROS)和脂质过氧化物(LPO)含量(P<0.05)。铜驯化对常温下ROS和LPO含量不产生影响,而显著降低了高温胁迫下大黄鱼ROS和LPO含量(P<0.05),表明铜驯化缓解了高温胁迫对大黄鱼的氧化损伤。从高温胁迫组vs对照组、铜驯化组vs对照组和(铜驯化+高温胁迫)组vs高温胁迫组中分别筛选到828个、2 311个和1 084个差异基因。GO和KEGG分析发现,差异基因主要富集在与TCA循环、氧化磷酸化、谷胱甘肽代谢、内质网蛋白加工、吞噬体和MAPK信号通路等相关功能上。聚类分析表明,高温胁迫下调了TCA循环、氧化磷酸化、谷胱甘肽代谢、吞噬体和MAPK信号等通路中的大部分基因表达,上调了内质网蛋白加工通路中的大部分基因表达,而铜驯化则对高温胁迫下大黄鱼的这些基因表... 相似文献
8.
斑马鱼(Danio rerio)早期胚胎发育囊胚时期是胚胎发育过程的关键时期,利用i TRAQ蛋白质质谱分析技术,检测斑马鱼早期胚胎发育过程中囊胚sphere时期的蛋白质表达情况,并分析该时期表达的蛋白质的相应功能和参与调控的生物过程。以野生型斑马鱼发育至sphere时期即4 hpf的胚胎为样本,利用i TRAQ标记与LC-MS/MS串联质谱技术,结合数据库比对,对该时期表达的蛋白质进行定性和定量的鉴定分析。检测结果共鉴定到的总蛋白数为1 178个,利用生物信息学进行功能分析,发现这些蛋白广泛参与了细胞信号传递、细胞运动和细胞骨架构建、细胞增殖、细胞分化、物质合成与代谢等各项重要的生命活动过程。研究表明,利用i TRAQ标记的方法可以对4hpf时期的斑马鱼胚胎中的蛋白质进行有效的分离和鉴定,并初步建立了斑马鱼早期发育关键阶段sphere时期的蛋白质组表达图谱,以期为斑马鱼胚胎发育过程中蛋白质组学和调控机制的研究提供参考。 相似文献
9.
大黄鱼冷诱导结合蛋白(CIRP)基因cDNA克隆及低温胁迫对其时空表达的影响 总被引：1，自引：0，他引：1
为研究冷诱导结合蛋白(CIRP)在大黄鱼低温适应过程中的作用,采用同源克隆技术获得了大黄鱼CIRP基因,并通过实时荧光定量PCR(qRT-PCR)技术检测了低温胁迫下大黄鱼各组织中CIRP基因的表达变化。结果显示,大黄鱼CIRP基因cDNA序列全长1211 bp,推测编码一个由182个氨基酸组成、分子量为19 ku的蛋白。序列比对显示硬骨鱼类CIRP基因序列较为保守,特别是N端的RNA识别基序(RRM)高度保守。系统进化树分析显示大黄鱼CIRP与银鳕相似性最高(81.5%),所有硬骨鱼类聚为一个大支。慢性低温胁迫中(12℃缓降至6℃),皮肤和肌肉中CIRP基因的表达量呈随温度降低而升高的趋势,6℃时表达量分别为12℃时的11.56倍和9.03倍;鳃、脑和心脏中CIRP基因表达量均呈先显著上升后显著下降的趋势,其峰值均出现在8℃时,表达量分别为12℃时的5.07、7.69和2.83倍;肠、肾脏、肝脏和脾脏中的表达量随温度降低而下降,6℃时的表达量最低,与12℃时相比降幅分别为80.0%、65.6%、91.2%、55.6%。急性低温胁迫(由12℃骤降至8℃并胁迫4 h)条件下,鳃、皮肤、脑、肝脏、心脏中CIRP基因表达量均呈先升高后降低的变化,反映了机体对低温胁迫的应激、适应过程;肌肉和肠中的表达量则始终显著低于胁迫前。慢性和急性低温胁迫中各组织CIRP表达量变化的差异提示,大黄鱼机体在低温胁迫响应和适应中有着多种调节机制。研究表明,CIRP基因参与了大黄鱼应对低温胁迫的过程。 相似文献
10.
为探究日本沼虾(Macrobrachium nipponense)应对高温胁迫的响应机制, 设置了水温对照组(20±0.5) ℃和高温组(30±0.5) ℃, 溶氧浓度(6.5±0.5) mg/L, 分别在高温胁迫 0 h、12 h、24 h、36 h、48 h 测定了日本沼虾肝胰腺和鳃组织中热休克蛋白 21 (heat shock protein 21, HSP21)、热休克蛋白 60 (heat shock protein 60, HSP60)、热休克同源蛋白 70-3 (heat shock protein cognate 70-3, HSC70-3)和热休克蛋白 90 (heat shock protein 90, HSP90)基因的表达, 相关抗氧化酶——超氧化物歧化酶(superoxide dismutase, SOD)、谷胱甘肽 S-转移酶(glutathione S-transferase, GST)、 谷胱甘肽过氧化物酶(glutathione peroxidase, GPX)和过氧化氢酶(catalase, CAT)活性以及组织结构的变化。结果表明, 高温胁迫下肝胰腺和鳃组织中的 4 个热休克蛋白基因均被诱导表达上升, 其中 HSC70-3 基因表达上升最为显著; 同时, 肝胰腺中的基因表达上升趋势比鳃中的基因表达趋势更为明显。高温胁迫下相关抗氧化酶活力均发生不同程度的变化, 肝胰腺中 SOD、GST 和 CAT 酶活力均有显著上升趋势, 而 GPX 酶活力仅有轻微变化; 鳃组织中的 SOD 酶活性显著低于对照组(P＜0.05), GST、GPX 和 CAT 酶活性显著高于对照组(P＜0.05)。肝胰腺和鳃组织结构均发生变化, 肝胰腺组织中分泌细胞及内部转运泡体积增大, 鳃组织结构层状上皮轻微弯曲, 血细胞排列紊乱等。综上所述, 高温胁迫激活了日本沼虾抗氧化系统, 并诱导了热休克蛋白基因的表达上升, 其中 HSC70-3 基因可能在日本沼虾的耐热性中起重要作用。高温胁迫下抗氧化酶活力的变化可能对于避免氧化损伤至关重要。本研究旨在通过了解日本沼虾应对高温胁迫的响应机制从而为日本沼虾的健康养殖提供参考。 相似文献
11.
Heat shock protein 70 expression in different tissues of Cirrhinus mrigala (Ham.) following heat stress 总被引：2，自引：0，他引：2
Members of heat shock protein 70 (HSP70) are highly conserved proteins of about 70 kDa and play important roles in protein folding. Levels of these proteins increase when cells are under stress. Environmental temperature influences both the basal and induced levels of HSPs. However, studies on HSPs in fishes from a tropical country such as India are lacking. In the present study, Indian major carp (IMC) Cirrhinus mrigala (Ham.) acclimatized at 25±2°C had high levels of HSP70, viz., 1.2–1.3 ng μg?1 total protein in kidney and gill and 4.2–5.3 ng μg?1 total protein in liver and brain tissues, indicating the presence of biochemically significant levels of stress. However, maintenance of acclimatized fish at 17°C for up to 48 h did not lead to a significant decrease in stress protein levels. A heat shock at 37°C for up to 48 h resulted in only two to threefold increase in HSP70 levels in these organs. Although the increase in HSP70 levels was apparent from the first hour of heat stress in all these tissues, the increase was significant from the second hour in the brain, the sixth hour in liver and kidney and the 20th hour in the gills. 相似文献
12.
为建立大黄鱼肿大细胞病毒的培养方法,明确其分类地位,用肿大细胞病毒检测呈阳性的大黄鱼幼鱼病料 (FD201807和SA201808)肾组织匀浆液感染鳜仔鱼细胞系 (mandarin fish fry cell line-1,MFF-1)并连续传代,从病料组织匀浆液和细胞冻融液中提取病毒DNA,克隆病毒主要衣壳蛋白基因 (mcp),测序后与NCBI GenBank中的虹彩病毒科肿大细胞病毒属病毒mcp以及2018—2020年所检出的15株大黄鱼肿大细胞病毒mcp进行比对分析。结果显示,病毒传至第4代才可引起MFF-1细胞病变,细胞病变的主要特征为细胞脱壁、变圆、折光度增强;感染时间越长脱壁细胞越多,同时培养液中的颗粒增加;透射电镜下可见感染细胞的细胞质散在大小为130~150 nm的六边形病毒粒子和空壳。感染细胞的病变周期随传代代次的增加而缩短,第15代次的FD201807株感染细胞80%细胞病变的时间为3 d,第15代次的SA201808株感染细胞80%细胞病变的时间为7~8 d。mcp序列比对和聚类分析发现,SA201808株与FD201807株的mcp序列存在21个碱基差异,二者的mcp序列分别与大黄鱼虹彩病毒(large yellow croaker iridovirus, LYCIV) LYCIV-Zhoushan (GenBank: MW139932.1)和花鲈虹彩病毒 (Lateolabrax maculatus iridovirus, LMIV) (GenBank: MH577517.1)相近。15株从大黄鱼病料检出的肿大细胞病毒中,12株的mcp序列与SA201808株聚类;3株与FD201807聚类。本研究利用MFF-1细胞系分离培养了大黄鱼肿大细胞病毒,揭示了大黄鱼肿大细胞病毒存在差异,为更好地了解大黄鱼肿大细胞病毒提供了数据参考。 相似文献
13.
Differential HSP90α expression in fish hepatocytes from polluted estuary during summer 总被引：2，自引：0，他引：2
The molecular chaperone, heat shock protein 90α (HSP90α), plays an important role in protein folding, degradation of denatured
proteins and steroid activation. It is essential for the maintenance of cellular integrity and survival when induced in response
to environmental, physical and chemical stresses. In the present investigation the effect of environmental stress on HSP90α
expression was examined in grey mullet Mugil cephalus living in either a contaminated (Ennore) or uncontaminated (Kovalam) estuary over two seasons: Hepatocytes were isolated
from grey mullet of both estuaries. Oxidative stress was determined along with HSP90α in these fish. Additionally, immunohistochemical
changes were studied to confirm the HSP90α expression. Comparison of the results revealed enhanced hepatocyte oxidative stress
and HSP90α expressions in fish from Ennore to a significant extent than fish from Kovalam. Also, the results showed significant
seasonal variations with maximum expression observed during summer compared to the monsoon season. Overexpression of HSP90α
in hepatocytes exposed to chronic environmental stress by pollutants may confer differential effects on cell survival by protecting
against oxidative stress induced changes. The results also indicate that seasonal variations have significant effect on the
HSP90α expression. 相似文献
14.
A. Rollo R. Sulpizio M. Nardi S. Silvi C. Orpianesi M. Caggiano A. Cresci O. Carnevali 《Fish physiology and biochemistry》2006,32(2):167-177
Two bacterial strains, Lactobacillus fructivorans, isolated from sea bream (Sparus aurata) gut, and Lactobacillus plantarum, isolated from human faeces, were administered simultaneously, during sea bream development, using Brachionus plicatilis and/or Artemia salina as vectors. The probiotic treatment significantly affected gut colonization. To test the probiotic influence on stress responsiveness, sea bream fry, 47 days post-hatching (p.h.), were subjected to pH stress (from 8.6 to 6.3) and cumulative mortality, cortisol levels and HSP70 gene expression were analysed. Cortisol was selected, since under stress conditions its level increases. HSP70 was selected with consideration of its wide involvement in response to a great number of injuries, and because it protects cells probably by binding and refolding damaged proteins. The results obtained indicated that the administration of probiotic to sea bream fry induced higher HSP70 levels, indicating a greater potentiality to respond to the harmful conditions possibly present in fish farms. This hypothesis is supported by the fact that the levels of cortisol found were significantly lower (P < 0.05) in both groups under probiotic treatment. When pH was used as a stressor, it induced a higher cumulative mortality in the control; the mortality was found to be significantly lower in both treated groups. Interestingly, a significant increase (P < 0.05) in HSP70 gene expression was observed in all stressed groups. These results suggest an improvement in tolerance to acute stress of fry fed with probiotics. 相似文献
15.
WD Repeat‐containing Protein 73, A Novel Gene Correlated with Gonad Development in Large Yellow Croaker,Larimichthys crocea 
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下载免费PDF全文 Dong L. Zhang Da H. Yu Hui Y. Luo Zhi Y. Wang 《Journal of the World Aquaculture Society》2016,47(2):268-276
The genome of Larimichthys crocea harbors more than 200 putative WD40 proteins. Yet, the function of these WD40 proteins remains unknown. Here a WD40 gene, wd73, was cloned and characterized in L. crocea (named as Lcwdr73). The cDNA is 1706 bp, including a 5′ untranslated region (UTR) of 51 bp, a 3′ UTR of 509 bp, and an open reading frame of 1146 bp encoding 381 amino acid residues. The putative LcWDR73 protein contains three WD40 domain motifs. The tissue expression profiles of both sexes revealed that Lcwdr73 was extremely highly expressed in testis and ovary, moderate in kidney and brain, and low in the other tissues by quantitative polymerase chain reaction analysis. Based on histology, the gonad development of L. crocea was divided into four phases: proliferative stage (Phase I), small developmental stage (Phase II), grand developmental stage (Phase III), and mature stage (Phase IV). With the gonad development, Lcwdr73 mRNA increased and reached a peak in Phase IV, and the expression in testes is significantly higher than that in ovaries, especially from Phase II to Phase IV, indicating that Lcwdr73 may play an important role during gonad development in L. crocea. The resutlts lay a foundation to control precocious puberty of L. crocea. 相似文献
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为比较日本鲭和大黄鱼肌肉中微生物和代谢功能的变化及其与鱼肉腐败特性之间的关系,本研究检测了2种鱼在冷藏过程中的理化指标和菌落总数的变化,利用Illumina Miseq测序技术分析细菌群落变化,并利用皮尔森相关性分析检验微生物与鱼肉腐败及组胺产生相关性,结合功能预测分析细菌群落组成与代谢功能之间的关系。结果显示,冷藏期间日本鲭和大黄鱼的pH、挥发性盐基氮、组胺、菌落总数等均呈上升趋势,且日本鲭上升较快;冷藏末期2种鱼TVB-N值和组胺含量分别达到76.34、59.98和59.92、3.11 mg/100 g;日本鲭肌肉中细菌丰富度和多样性先增加后减少,大黄鱼则整体呈下降趋势;2种鱼肌肉中的优势腐败菌均为希瓦氏菌属;日本鲭体内与TVB-N产生相关的菌共12种,其中10种与组胺产生具有显著相关性;大黄鱼体内与TVB-N产生相关的菌共7种,但未检测出与组胺产生具有相关性的细菌;冷藏过程中氨基酸代谢和碳水化合物代谢为最主要的代谢通路,日本鲭样品组氨酸、精氨酸、脯氨酸等氨基酸代谢相关基因和丁酸丁酯代谢、丙酸酯代谢及丙酮酸代谢丰度均显著高于同一时期的大黄鱼,本实验从微生物代谢水平解释了日本鲭比大黄鱼更易腐败的原因,为不同水产品腐败特性的研究提供新思路。 相似文献
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D. Jirsa D.‐F. Deng D.A. Davis W.‐F. Wang S.S.O. Hung M. Drawbridge 《Aquaculture Nutrition》2013,19(2):227-232
A feeding trial was conducted to study the effect of dietary lipid on growth performance and heat‐shock protein (HSP70 and HSP60) response of white seabass (WSB), Atractoscion nobilis. Five diets were formulated to contain 440 g kg?1 protein from 300 g kg?1 fish meal, 240 g kg?1 soybean meal and 100 g kg?1 soy protein concentrate with different levels of lipid: 100, 120, 140, 160 or 180 g kg?1. At the end of the trial, heat shock response based on HSP70 and HSP60 was measured in liver and white muscle from fish at ambient temperature and temperature shock conditions. Final weight and percent gain were significantly higher for fish fed the 100 g kg?1 lipid diet than for fish fed the rest of the diets (P ≤ 0.05). Feed conversion ratio was lowest for fish fed the 100 g kg?1 lipid diet. The HSP70 and HSP60 responses were positively correlated to dietary lipid levels following temperature shock. At ambient temperature, HSP60 and HSP70 responses in muscle and HSP60 response in liver increased with dietary lipid level. Temperature shock significantly increased the HSP response of fish in all treatments. Results of this study demonstrated that a moderate (110–120 g kg?1) level of dietary lipids would be recommended for production diets but a higher dietary lipid level may be required for optimal stress tolerance. 相似文献