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1.
对三角鲂(mEGALOBRAMA TERMINALIS)形态学特征进行研究，用RT—PCR的方法从三角鲂肝脏克隆了三角鲂胰岛素样生长因子-Ⅰ(IGF-Ⅰ)基因的cDNA序列。三角鲂传统的形态学数据与团头鲂相似。序列分析表明，三角鲂ICE-Ⅰ cDNA由486个核苷酸构成，编码161个氨基酸，包含整个信号肽、B、C、A、D和E区，与鲂属团头鲂比较，三角鲂与团头鲂IGF-Ⅰ cDNA序列同源性为99．8％，氨基酸序列同源性为99．4％。由此可见三角鲂的遗传学特征与团头鲂非常相似。 相似文献
2.
The insulin-like growth factor I (IGF-I) cDNA (GenBank Accession No. AY247412) of triangular bream (Megalobrama terminalis) was expressed for the first time in Escherichia coli. To construct the expression plasmid, the IGF-I cDNA was subcloned into prokaryotic-expressing vector pGEX-4T-1. The E.coli JM109 was transformed with the recombinant plasmid pGEX-4T-1-IGF-I, and the transgene expression was observed after being induced with isopropyl-β-D-thiogalactoside (IPTG). The results of SDS polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting indicated that the recombinant fusion protein had immune activity and the molecular weight was about 47 kDa. The results of SDS-PAGE and thin layer scanning showed that the yield of fusion protein had been enlarged with prolonging time. When the time of induced expression was 1, 2, 3, 4, 5, and 6 h, the expression amount was approximately 1.4, 4.3, 8.1, 11.3, 16.3 and 18.8% of total bacterial protein respectively. 相似文献
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S. He Z. Wu Y. Liu N. Wu Y. Tao L. Xu Z. Zhou B. Yao E. Ring 《Aquaculture Nutrition》2013,19(5):765-772
Dried distiller's grain (DDG) is considered as an alternative ingredient of dietary feed due to its high contents of protein, fibre and fat. In this study, 60 g kg‐1 of DDG was used to feed grass carp (Ctenopharyngodon idella), bluntnose black bream (Megalobrama amblycephala), gibel carp (Carassius gibelio) and black carp (Mylopharyngodon piceus) for 8 weeks, and its effect on fish production and gut allochthonous microbiota was investigated for the development of a suitable fish feed high in nutrients and low in cost for polyculture freshwater fish. DDG supplementation resulted in the less weight gain and higher feed conversion ratio of black carp (P < 0.05), but had no significant effects on other fish or parameters. PCR–denaturing gradient gel electrophoresis (DGGE) analysis indicated that all four fish species had some common and unique bacteria in their digestive tracts, and the gut microbiota of bluntnose black and gibel carp fed the control diet and DDG diets were very similar (Cs > 91%); of them, the total counts of intestinal bacteria studied by qPCR increased in grass carp (P < 0.05) and depressed in black carp (P < 0.05) when fed dietary DDG. Thus, we assumed that dietary DDG modulated production and gut microbiota of fish in a host‐specific way. 相似文献
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为探究Tyrosinase(Tyr)基因在黄河鲤体色形成中的作用,利用生物显微镜对黄河鲤黑色鳞片、银色鳞片、鳍条色素细胞的组成进行观察,并利用RACE克隆黄河鲤Tyr基因全长cDNA序列,荧光定量PCR分析其在不同组织中的表达情况,Western blot印迹和免疫组织化学方法检测其蛋白的表达定位。结果显示,黄河鲤具有黑色素细胞、黄色素细胞和虹彩细胞3种色素细胞,3种色素细胞的数量、种类和分布在不同组织中存在较大差异。黄河鲤Tyr基因cDNA全长1717 bp(GenBank ID:No.KY305667),其中ORF长1605 bp,编码535个氨基酸,5′-UTR区41 bp,3′-UTR区71 bp。蛋白同源分析发现,黄河鲤Tyr与鲤科鱼类相似性最高,与哺乳类及鸟类等脊椎动物相似性最低。系统进化分析显示,黄河鲤Tyr与鲤、瓯江彩鲤、锦鲤和斑马鱼等鲤科鱼类的亲缘关系最近。实时定量PCR分析表明,Tyr基因在黄河鲤不同组织中均有表达,肌肉中的表达量最高,其次是黑色皮肤,另外在黄色皮肤、臀鳍和尾鳍等黑色素细胞存在的组织中也有较高表达。Western blot印迹结果显示,Tyr基因在黑色皮肤中表达最高,其次是臀鳍和尾鳍,银色皮肤中没有表达。免疫组织化学结果显示,Tyr基因在黑色皮肤的黑色素细胞和黏液细胞中有表达。研究表明,Tyr基因表达水平与黑色素细胞数量和分布具有一定相关性,参与黄河鲤体色的形成。 相似文献
6.
Comparative study on the effect of high dietary carbohydrate on the growth performance,body composition,serum physiological responses and hepatic antioxidant abilities in Wuchang bream (Megalobrama amblycephala) and black carp (Mylopharyngodon piceus Richardson, 1846) 
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下载免费PDF全文 Chuanpeng Zhou Xianping Ge Bo Liu Jun Xie Ruli Chen Linghong Miao Mingchun Ren 《Aquaculture Research》2017,48(3):1020-1030
A 90‐day trial investigated the effect of dietary carbohydrate level on growth performance, body composition, serum physiological responses and hepatic antioxidant abilities of a herbivorous fish, Wuchang bream (Megalobrama amblycephala) and a carnivorous fish, black carp (Mylopharyngodon piceus Richardson, 1846). For each species, two isonitrogenous and isolipidic semi‐purified diets (28.6% crude protein and 5.3% crude lipid for Wuchang bream) were formulated to contain 30.4% and 52.9% carbohydrate (WB‐NCHO and WB‐HCHO); Two isonitrogenous and isolipidic semi‐purified diets (32.6% crude protein and 5.3% crude lipid for black carp) were formulated to contain 20.5% and 40.5% carbohydrate (BC‐NCHO and BC‐HCHO). It was shown that, compared to Group WB‐NCHO, Wuchang bream in Group WB‐HCHO had lower weight gain rate, specific growth rate, serum total protein, hepatic superoxide dismutase activity and total antioxidative capacity, while higher feed conversion ratio, serum triglyceride and cortisol levels (P < 0.05). Compared to Group BC‐NCHO, black carp in Group BC‐HCHO had lower final weight, weight gain rate, specific growth rate, protein efficiency ratio, serum alkaline alkaline phosphatase activity, serum total protein, hepatic superoxide dismutase activity and total antioxidative capacity, while higher feed conversion ratio, whole body crude lipid, serum triglyceride, cholesterol, glutamic‐oxaloacetic transaminase activity and cortisol levels (P < 0.05). The results of this study suggested that Wuchang bream had better capacity of utilizing dietary carbohydrate than that of black carp, while high levels of dietary carbohydrate might not only be detrimental to growth performance and feed utilization but also the immune response and health for both species. 相似文献
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为研究草鱼抗菌肽NK-lysin基因的组成结构、在组织中的表达差异及其抑菌活性,实验根据已知鱼类NK-lysin基因保守区序列设计上下游引物,采用RT-PCR和RACEPCR技术克隆草鱼NK-lysin基因(Cinkl),RT-PCR进行各组织间的表达分析,并构建Cinkl成熟肽的原核表达载体,采用琼脂糖孔穴扩散法检测重组蛋白的抑菌活性。结果显示,Cinkl c DNA全长768 bp,编码121个氨基酸;基因组DNA全长3 361 bp,包含4个外显子和3个内含子。经多序列比对分析和结构域预测表明,Ci Nkl氨基酸序列具有SAPLIP家族的特征:含有6个保守的半胱氨酸和saposin B蛋白结构域,与斑马鱼Nklc、Nkld的相似性分别为57.98%和63.03%。构建NK-lysin氨基酸序列系统进化树,发现Ci Nkl与斑马鱼Nklc、Nkld聚为一支。RT-PCR检测结果显示,Cinkl m RNA在所有检测的草鱼组织中均有表达,在脾脏中表达量最高,心脏、鳃、中肾和头肾的表达量次之,皮肤、脑、肝脏和肠有微量表达。p ET-28b-MBP与Ci Nkl成熟肽序列构建的原核表达质粒在宿主菌Rosetta(DE3)表达出约57 ku的可溶性融合蛋白,利用Ni-NTA His·Bind树脂进行纯化,Ci Nkl重组蛋白对G–细菌如大肠杆菌M15株、嗜水气单胞菌,以及G+细菌如金黄色葡萄球菌均具有抑菌活性。基于草鱼NK-lysin的组织表达特征和重组蛋白的抑菌活性,推测NK-lysin在草鱼的先天性免疫防御中发挥调节作用。 相似文献
8.
Wenwen Wang Zhigang Zhou Suxu He Yuchun Liu Yanan Cao Pengjun Shi Bin Yao Einar Ringø 《Aquaculture Research》2010,41(9):e72-e83
PCR‐denaturing gradient gel electrophoresis (DGGE) was applied to analyse the microbial community attached to the gills and skin of poly‐cultured gibel carp (Carassius auratus gibelio) and bluntnose black bream (Megalobrama amblycephala Yih) and compare these results with those detected in the rearing water. The microbiota discussed included bacteria, fungi and a specific bacterial taxa of actinomycetes was also analysed. Proteobacteria, Firmicutes, Actinobacteria, Cyanobacteria, Ascomycota, Basidiomycota and some unclassified microbiota were identified. Based on our results, we concluded that: (1) the adherent bacterial/fungal communities on the gills and skin were different from those in the rearing water, (2) the bacterial/fungal diversities on fish gills were lower than that on fish skin, (3) the adherent bacterial/fungal communities on gill and skin of gibel carp were different from that of bluntnose black bream and (4) the adherent actinomycetal community showed certain similarity between the skin of different hosts. Based on our conclusions, we suggested that the topic investigated in the present study merits further investigations. 相似文献
9.
Interploid hybrids were produced by mating allo‐tetraploid males, an interspecific tetraploidy induced by blunt snout bream (Megalobrama amblycephala) ♀× black bream (Megalobrama terminalis) ♂, with blunt snout bream females. The fertilization rate was high in the interploid hybrids. Although significant (P<0.05) mortalities were observed at hatching, swim‐up and 30‐day stages, respectively, the survival percentage of the interploid hybrids became stabilized after the 60‐day stage. In different interploid hybridization batches, 64–97% of juveniles were identified as 3n and 1–36% as 2n. Less than 2% of tetraploids were obtained in these interploid hybridization batches. The daily growth rate of the interploid 3n increased 7.5%, compared with that of diploid blunt snout breams during the 180‐day culture. Moreover, a method using seven morphological parameter ratios was set up with a relatively high classification accuracy of 93.8% to harmlessly discriminate interploid 3n from allo‐tetraploids, black bream and blunt snout bream. 相似文献
10.
草鱼胰岛素样生长因子_基因克隆及序列分析 总被引：3，自引：1，他引：3
采用逆转录－聚合酶链式反应（ＲＴ－ＰＣＲ）方法,从草鱼肝脏的总ＲＮＡ中扩增出胰岛素样生长因子－Ｉ（ＩＧＦ－Ｉ）基因序列,定向克隆至质粒pUC18,测宇了该基因序列,推导期编码的蛋白序列,克隆的cDNA序列编码包括Ｂ,Ｃ,Ａ,Ｄ和Ｅ五个区域的１７个氨基酸,与鲤ＩＧＦ－Ｉ成熟肽比较,核酸序列和氨基酸序列的同源性分别为９３．８％和９７．１％,E区域分析结果表明,所克隆的草鱼ＩＧＦ－Ｉ序列属于ＩＧＦ－ＩＥa-2亚型。 相似文献
11.
利用One-Step RT-PCR技术,从鲫(Carassius auratus)肝胰脏组织中克隆了胰岛素样生长因子-Ⅰ(IGF-Ⅰ),并进行了序列分析。结果表明,鲫IGF-ⅠcDNA由486个核苷酸组成,编码包括信号肽、成熟肽(B、C、A、D四个域)和E域的161个氨基酸。核苷酸同源性分析揭示,鲫IGF-ⅠcDNA序列与金鱼的同源性最高,为99.18%;与鲮、三角鲂、团头鲂的同源性次之,分别为94.44%、94.65%、94.44%;与鲤、草鱼的同源性相对较低,为86.03%和85.29%。E区域的分析结果表明,鲫IGF-Ⅰ序列为Ea-2亚型。 相似文献
12.
Insulin-like growth factors I and II (IGF-I and IGF-II) are two highly homologous mitogenic peptides that are expressed ubiquitously and show diverse effects on development, growth, and metabolism. The cDNA encoding IGF-I of a teleost, the orange spotted grouper (Epinephelus coioides) was produced from liver by RT-PCR, and rapid amplification of cDNA ends, RACE. Typically, the deduced 186 amino acid protein contains a signal peptide, B, C, A, D and E domains. On the amino acid level, grouper IGF-I shares 97.3% similarity with black seabream (Sparus macrocephalus) with the differences focusing on the B and C domains. The analysis of the E domain showed that grouper IGF-I belonged to Ea-4 type. When mature amino acid sequence was compared with other vertebrates, it revealed higher similarity with black seabream and halibut, while lower similarity with human and mouse. The expression of IGF-I mRNA in adult tissues was studied using RT-PCR. IGF-I mRNA expression level in the liver was significantly higher than those in the brain and muscles. In other tissues, low amount of IGF-I mRNA expression was also detected. The coding region of IGF-I cDNA for mature IGF-I protein was subcloned into an expression plasmid pTRX and fused with E. coli thioredoxin (Trx). Moreover, we have successfully developed an expression system in E. coli to overproduce recombinant grouper IGF-I. Using western blotting, we found that the fusion protein could blot with antiserum to barramundi IGF-I further confirming the immunoactivity of the recombinant IGF-I. 相似文献
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D.R. Tocher G. Mourente A. Van der Eecken J.O. Evjemo E. Diaz M. Wille J.G. Bell Y. Olsen 《Aquaculture International》2003,11(1-2):195-216
The aim of the study was to compare theantioxidant systems in juvenile marine fish ofcommercial importance to European aquaculture,namely turbot (Scophthalmus maximus),halibut (Hippoglossus hippoglossus) andgilthead sea bream (Sparus aurata). Thepresent dietary trial was specifically designedto investigate the antioxidant effects ofvitamin E under moderate oxidising conditions,including high dietary levels of highlyunsaturated fatty acids and the feeding ofoxidised oils. The objective was to induce astressful pro-oxidant status to enablecharacterisation of the biochemical responsesto peroxidative stress without causingunnecessary suffering to the experimentalanimals or high mortalities during the trials. Both sea bream and turbot showed excellentgrowth, whereas growth was poorer in halibut.Dietary oxidised oil significantly reducedgrowth in turbot and especially in halibut, butnot in sea bream. Vitamin E improved growth insea bream fed oxidised oil but not in turbot orhalibut. However, vitamin E supplementationappeared to improve survival in all threespecies. In sea bream and turbot, liverantioxidant defence enzyme activities weregenerally increased by feeding peroxidised oiland reduced by vitamin E. Conversely, inhalibut, the liver antioxidant defence enzymeactivities were not increased by feedingperoxidised oil and only superoxide dismutasewas reduced by feeding vitamin E. Consistentwith these data, feeding oxidised oil increasedlipid peroxidation products in halibut, butgenerally not in sea bream or turbot.Furthermore, lipid peroxidation products weregenerally reduced by dietary vitamin E in bothsea bream and turbot, but not in halibut. Therefore, halibut liver antioxidant defenceenzymes did not respond to dietary oxidised oilor vitamin E as occurred in turbot and,especially sea bream. This resulted inincreased levels of lipid peroxides in halibutcompared to turbot and sea bream in fish givendietary oxidised oil. In addition, supplementalvitamin E did not reduce lipid peroxides inhalibut as it did in turbot and sea bream. Theincreased peroxidation stress in halibut mayaccount for their poorer growth and survival incomparison to turbot and especially sea bream.Halibut were reared at a lower temperature,although relatively high for halibut, thaneither turbot or sea bream but they were alsoslightly younger/smaller fish and possibly,therefore, more developmentally immature, andeither or all of these factors may be importantin the lack of response of the liver enzymes inhalibut. 相似文献
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本文旨在克隆鲤(Cyprinus carpio)激素敏感性脂肪酶a (hormone-sensitive lipase a, lipea)基因,探究其时空表达特征及其与脂肪沉积的关系,以期为鲤脂肪沉积分子调控机制研究奠定理论基础。本实验采用RACE技术克隆获得鲤lipea基因,其cDNA全长为3379 bp,包括230 bp的5′非翻译区(5′-UTR)、1067 bp的3′非翻译区(3′-UTR)和编码693个氨基酸的开放阅读框。多序列比对显示,鲤lipea与斑马鱼(Danio rerio)、牙鲆(Paralichthys olivaceus)和虹鳟(Oncorhynchusmykiss)的氨基酸序列相似性分别为88.17%、64.94%和63.35%,不同物种间序列差异主要集中在C-端的多肽序列上。蛋白质结构分析显示该蛋白含有3个功能域分别具有脂肪酶活性、乙酰基水解酶活性和水解酶活性。应用实时荧光定量PCR技术分析了lipea基因在鲤各组织、不同发育时期以及脂肪含量极端差异个体的肌肉和脂肪组织中mRNA表达量的变化。结果显示, lipea在各组织中均有表达,腹腔脂肪中相对表达量最高,其次是腹部肌肉,血液中最少。随着胚胎的发育, lipea的表达水平呈现下降趋势, 8细胞期表达量最高,其次是囊胚晚期,开口期最少。lipea表达量与背部肌肉脂肪含量呈正相关,但差异不显著(P0.05),与腹腔脂肪量呈负相关,且差异极显著性(P0.01),推测lipea基因对鲤腹腔脂肪沉积可能具有抑制作用。本实验结果可为进一步解析鲤lipea基因的功能和基因调控脂肪沉积分子机制研究提供参考。 相似文献
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为探究SPR在锦鲤体色形成中的作用,实验利用RACE技术获得spr cDNA全长序列,并分析其时空表达模式,同时利用Western blot和免疫组织化学方法检测SPR蛋白在皮肤、鳍条和鳞片中的分布和表达情况。结果显示,spr cDNA全长879 bp,包含132 bp和134 bp的5′和3′非编码区,开放阅读框510 bp,编码170个氨基酸残基。氨基酸序列比对和系统进化树分析显示,锦鲤SPR具有保守的adh_short_C2结构域,与金鱼相似性高达97.7%。spr在各组织中均有表达,其中皮肤的表达量最高。spr在锦鲤个体发育的4个阶段表现为先降后升。纯红、纯白及红白3种体色锦鲤皮肤、鳞片和鳍条中spr mRNA和蛋白表达水平基本一致,在纯红锦鲤皮肤中表达量最高,红白锦鲤白色皮肤、鳞片和鳍条的表达量最低。SPR组织定位分析显示,红色锦鲤和白色锦鲤皮肤中均检测到阳性信号,其中红色皮肤阳性信号强度高于白色皮肤。研究表明,spr可能与锦鲤红/黄色素细胞的分化和形成具有一定相关性,参与了锦鲤体色的形成。 相似文献
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Xinhong Guo Jinpeng Yan Shaojun Liu Bing Xiang Yun Liu 《Fish physiology and biochemistry》2010,36(2):125-133
To investigate the evolutional significance of Sox9 in fish, we isolated and characterized Sox9a cDNA and genomic clones in triploid crucian carp. The cDNA encoded a protein of 457 amino acids with an HMG box and showed
more than 60% amino acid sequence identity with known vertebrate Sox9 proteins. Triploid crucian carp and vertebrate Sox9s showed similar gene structure, and two introns in the coding region were located at conserved positions. On the basis of
the amino acid sequences, Sox9a can be categorized into the same subgroup of Sox-E proteins as Sox8, 9, and 10. Interestingly, the expression of triploid crucian carp Sox9a was predominantly observed not in the ovary but in the testis by Northern blot and RT-PCR analysis. The expression analysis
of Sox9a suggested that it may seldom contribute to the formation of normal functions of spermatozoa, but it may play an important
role in the development of testicular tubules. Besides the testicular expression, Sox9a was also shown to be expressed in many other tissues including the brain, kidney, and heart of triploid crucian carp, indicating
that Sox9 may have unique functions in some specific tissues during development. 相似文献
17.
Dapeng Li Jia Teng Hang Wang Xiaochang Liu Huiyi Wang 《Journal Of Aquatic Food Product Technology》2017,26(4):468-478
The aim of this study was to investigate the relationship between lipid oxidation, protein function properties, and freshness changes of blunt-snout bream (Megalobrama amblycephala) fillets treated with 2% and 4% salt during storage at 4°C. Salting with 2% and 4% salt could delay quality deterioration and protein denaturation, thus improving sensory attributes to some extent. But, 4% salt promoted lipid oxidation of blunt-snout bream fillets. There is a significant correlation (p < 0.05) between freshness indexes and lipid oxidation or protein function properties (total SH content, Ca2+-ATPase activity). Salting with 2% salt is an ideal treatment to control the quality of blunt-snout bream fillets stored at 4°C. 相似文献
18.
Reciprocal hybrids between tench Tinca tinca (L.) and carp Cyprinus carpio L., and tench and bream Abramis brama (L.) were produced artificially. The survival of all these hybrids during embryogenesis was quite high. The highest survival
rate (over 60%) at the eyed stage was observed for tench and bream hybrids (both sex combinations). The hatching rates of
these hybrids were also over 60%. The number of larvae with some abnormalities (i.e. deformed body) was low. In contrast,
the hatching rates of tench and carp hybrids were very low (0.2%). From over 1000 fertilized eggs, only three specimens started
swimming, and only one specimen survived to juvenile stage. Embryos of hybrids and their parental species differ in morphological
features. These differences were also visible in the juveniles. Body parameters of juvenile hybrids produced from three species
had intermediate values in comparison to parental fish. 相似文献
19.
Marek Matras Magdalena Stachnik Ewa Borzym Joanna Maj‐Paluch Michal Reichert 《Journal of fish diseases》2019,42(7):959-964
During a PCR‐based CEV survey in Poland in 2015–2017, the virus was detected in many farms both in clinical and asymptomatic cases and in common as well as in koi carp (Cyprinus carpio). In order to evaluate the potential carrier role of fish species that share the same habitats with carp, an experimental trial was performed. Investigations carried out on specimens of bleak (Alburnus alburnus), crucian carp (Carassius carassius), European perch (Perca fluviatilis), Prussian carp (Carassius gibelio), roach (Rutilus rutilus) and tench (Tinca tinca) cohabited with CEV‐infected carp yielded positive results. These species of fish were experimentally cohabited with CEV‐infected common carp at a temperature of 16°C ± 1. Material from the brain, gills, spleen, kidneys, intestine and skin was investigated for the presence of CEV DNA. Similar investigations were performed with uninfected fish designated controls. Samples were tested for CEV by qPCR. 相似文献
20.
为研究Akt基因在中华绒螯蟹蜕壳前后肌肉生长过程中的功能,应用RACE技术克隆得到编码中华绒螯蟹Akt(命名为Es Akt)的全长为2 200 bp的c DNA序列,包括159 bp的5′非翻译区(5′-UTR)、496 bp的3′非翻译区(3′-UTR)和长度为1545 bp编码514个氨基酸的编码序列。蛋白质结构域分析显示Es Akt含有丝氨酸/苏氨酸蛋白激酶家族的3个特征性保守结构域。多序列比对和系统发育分析显示,Es Akt与中国明对虾、凡纳滨对虾的Akt序列一致性都为0.889,在系统发育树中节肢动物Akt形成一个分支。应用荧光定量RTPCR技术分析Es Akt在性成熟中华绒螯蟹各组织及幼体不同蜕壳时期不同部位肌肉组织中转录水平上表达量的变化。结果显示,Es Akt在性成熟个体的肝胰腺、眼柄、表皮、卵巢、精巢、心脏、螯足、鳃、三角膜等组织中均有表达,其中卵巢、眼柄和精巢中表达量较高,肝胰腺中表达量最低。在幼体不同蜕壳时期不同部位的肌肉中,Es Akt表达量变化不同,步行足肌肉组织中Es Akt m RNA无显著的统计学差异。腹部肌肉组织中Es Akt m RNA水平在蜕壳前晚期D3~D4期表达量显著高于蜕壳后A~B期和蜕皮间期C期。螯足肌肉在蜕壳前晚期D3~D4期急剧下调,蜕壳后A~B期开始表达量显著升高,直至蜕皮间期C期。研究表明,Es Akt在中华绒螯蟹蜕壳过程中不同部位肌肉组织中的表达量变化与蜕壳周期密切相关,说明Es Akt参与中华绒螯蟹蜕壳诱导的肌肉萎缩、生长及重建过程。 相似文献