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1.
采用RACE技术,克隆出三疣梭子蟹(Portunus trituberculatus)核受体基因HR38的c DNA全长,命名为PTHR38。该基因c DNA序列全长为2950 bp,5′和3′非编码区(UTR)长分别为101 bp和551 bp,开放阅读框(ORF)长2298 bp。推测编码765个氨基酸,预测分子量为84.2 k D,理论等电点为6.3。生物学分析预测,PTHR38基因编码的蛋白属于不稳定蛋白,不具有跨膜结构域。同源性分析表明,PTHR38基因编码的氨基酸序列与大型溞(Daphnia magna)的同源性最高。实时荧光定量分析表明,PTHR38基因在蜕皮周期的各个组织中均有差异表达,在眼柄中表达差异最大。去除单侧眼柄后,PTHR38的表达量在整体上呈现不同程度的上升趋势。 相似文献
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实验克隆了兴国红鲤(Cyprinus carpio var singuonensis)雄激素受体(androgen receptor,AR)基因的全长c DNA序列和卵黄蛋白原(vitellogenin,vtg)基因的部分c DNA序列,并对雌性个体组织及甲基睾丸酮(Methyltestosterone,MT)暴露下幼鱼肝胰腺(以下简称:肝)中AR和vtg的表达进行检测。兴国红鲤AR基因c DNA全长3 229 bp,包括104 bp的5'非编码区(untranslation region,UTR)、编码846个氨基酸的2 541 bp开放阅读框(open reading frame,ORF)和485 bp 3'UTR。氨基酸序列同源性分析表明,兴国红鲤AR与其他鲤科鱼类AR的同源性较高。组织表达特征研究表明,AR在雌性个体的肝、卵巢、中肾、肌肉和端脑中有表达,其中肝中的表达量最高;vtg主要在肝、端脑和鳃中表达。兴国红鲤幼鱼在50μg/L的MT中暴露4周后,采用qRT-PCR检测了肝中AR和vtg的表达情况。在处理第1、2周,AR的表达受到一定的抑制,而在第3、4周其表达量升高,但其表达无显著性变化;而vtg的表达量在第3、4周均有极显著的升高。 相似文献
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利用RT-PCR和RACE方法克隆得到镜鲤Cyprinus carpio肝脏中高不饱和脂肪酸(HUFA)合成代谢的脂肪酸去饱和酶(△6FAD)基因c DNA全序列。结果表明:镜鲤△6FAD基因c DNA全长为1 446bp,开放阅读框(ORF)为1 332bp,编码444个氨基酸。编码的蛋白序列包含FAD全部特征结构区,包括1个细胞色素b5结构域、2个跨膜区和3个组氨酸簇,与其他鱼类的Δ6FAD氨基酸序列具有69.0%~92.0%的同源性。系统树分析显示:与斑马鱼Danio rerio的亲缘关系最近。实时荧光定量PCR(RT-q PCR)检测发现:脂肪酸延长酶基因在镜鲤肝脏中表达量最高,背部肌次之,在血液中表达最低。本研究结果为进一步研究镜鲤HUFA的合成途径及调控机理提供了基础资料。 相似文献
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利用cDNA末端快速扩增法(RACE)克隆团头鲂Megalobrama amblycephala铁蛋白基因c DNA全长序列;同时研究经嗜水气单胞菌Aeromonas hydrophila攻毒后团头鲂肝组织中铁蛋白表达的变化,了解铁蛋白基因在团头鲂免疫应答中的作用。结果表明:团头鲂铁蛋白c DNA全长序列包括150bp的5’末端序列(untranslated region,UTR),270bp的3’UTR,以及522bp编码174个氨基酸的开放阅读框(open reading frame,ORF)。这些氨基酸序列同其他鱼类铁蛋白M链氨基酸序列同源性较高。团头鲂铁蛋白基因在5’非编码区核甘酸序列124~154的位置有个特殊的结构,即铁反应元件(iron response element,IRE)。荧光定量PCR分析表明:铁蛋白基因在团头鲂肌肉、心脏、鳃、肝胰脏、脑和肾脏等组织器官中都有表达,在肝胰脏的表达量最高,脑组织表达量最低。经嗜水气单胞菌急性感染后,团头鲂肝胰脏组织中铁蛋白基因表达量显著上调。上述结果表明:团头鲂铁蛋白M亚基在团头鲂免疫应答过程中起重要作用。 相似文献
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为在分子水平解析牙鲆(Paralichthys olivaceus)抗淋巴囊肿病的机理,本研究克隆牙鲆淋巴囊肿抗病免疫相关基因efhd2和tbc1d25的c DNA全序列,运用生物信息学方法对基因序列进行了详细分析;同时,采用荧光定量PCR分析了efhd2和tbc1d25基因在牙鲆胚胎发育不同阶段、淋巴囊肿抗病和患病个体不同组织的表达特征。结果显示,efhd2基因c DNA全长为5231 bp,开放阅读框(ORF)长为699 bp,编码232个氨基酸。tbc1d25基因c DNA全长为3173 bp,ORF长为2601 bp,编码866个氨基酸。定量结果显示,efhd2和tbc1d25基因在胚胎发育的各个时期均有不同程度的表达,其中,efhd2在出膜仔鱼期表达量最高,而tbc1d25在受精卵中的表达量显著高于其他时期(P0.05)。在所研究的淋巴囊肿抗病和患病个体不同组织中,efhd2和tbc1d25基因均有不同程度的表达,抗病个体的血液中,这2个基因的表达量均显著高于患病个体(P0.05)。本研究结果为深入探讨efhd2和tbc1d25基因功能和解析牙鲆淋巴囊肿抗病机理提供了基础资料。 相似文献
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旨在研究异齿裂腹鱼(Schizothorax o’connori)缩胆囊素(cholecystokinin, CCK)基因的摄食功能。本研究克隆得到了异齿裂腹鱼CCK基因的cDNA全长,通过生物信息学分析发现其属于CCK-1亚型。异齿裂腹鱼CCK的cDNA全长为773bp,其中开放阅读框(ORF)为372bp,可以编码123个氨基酸。异齿裂腹鱼CCK由1个信号肽和1个典型的CCK-8肽保守结构域组成,为亲水性蛋白,但没有跨膜结构。运用实时荧光定量PCR(Real-timePCR)检测异齿裂腹鱼CCK基因在组织中的分布情况,以及餐前餐后和禁食复喂对其表达量的影响。结果表明,异齿裂腹鱼CCK在各组织中均有表达,其中在脑中表达量最高,在肠道、心脏、肝、脾、肾、皮肤、鳃、眼和鳔中表达量相对较高,在肌肉中表达量最低。餐后CCK基因的表达量显著升高,禁食使异齿裂腹鱼CCK基因的表达量显著下降,而复喂使CCK基因的表达量显著上升,表明CCK基因既是异齿裂腹鱼的餐后饱感信号因子,又是长期调控摄食因子。本研究为异齿裂腹鱼的人工饲养和品种保护等提供了理论依据。 相似文献
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通过cDNA末端快速扩增(Rapid amplification of c DNA ends,RACE)技术克隆得到魁蚶(Scapharca broughtonii)C型凝集素(C-type lectin,Sb-Lec1)基因,该基因全长为700 bp,其中,5′-UTR为29 bp,3′-UTR为167 bp,开放阅读框长度为504 bp,编码167个氨基酸,包括长度为23个氨基酸的信号肽序列、129个氨基酸的糖识别结构域(CRD)以及参与二硫键形成的6个半胱氨酸。预测蛋白分子量为19.11 k Da,理论等电点为4.74。多序列比对结果显示,Sb-Lec1基因CRD编码的氨基酸序列与长牡蛎(Crassostrea gigas)、紫贻贝(Mytilus galloprovincialis)和海湾扇贝(Argopecten irradians)C型凝集素的同源性分别为38%~40%、34%~35%和38%~39%,Sb-Lec1基因编码的氨基酸序列与其他物种的凝集素基因具有相似的结构,均含有形成二硫键的4个保守半胱氨酸。系统进化分析结果显示,魁蚶先与贝类聚为一支,再与脊椎动物聚在一起,表明魁蚶Sb-Lec1基因在进化树上的位置与其传统分类所处位置一致。采用荧光定量PCR技术,检测了Sb-Lec1基因在组织中的表达情况,发现其在肝胰腺、血淋巴、鳃、外套膜、闭壳肌、斧足中均有表达,其中,肝胰腺表达量最高。同时,分析了Sb-Lec1基因在鳗弧菌(Vibrio anguillarum)刺激下的mRNA表达量变化情况。结果显示,与对照组相比,菌刺激组Sb-Lec1基因mRNA在各检测组织中的表达量均显著上调(P0.05),随着刺激时间的延长,表达量呈先升高后降低的趋势。本研究表明,魁蚶Sb-Lec1基因在机体免疫防御方面发挥重要功能。 相似文献
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仿刺参EGFR基因的克隆与表达分析 总被引：1，自引：1，他引：0
表皮生长因子受体(EGFR)是多种细胞因子的受体,在细胞增殖、迁移及分化中具有重要的作用。应用RACE法首次从仿刺参体腔细胞中克隆出EGFR基因的全长cDNA序列。该cDNA全长3 826 bp,包括821 bp的5’-UTR,281 bp的3’-UTR,开放阅读框2 724 bp,编码907个氨基酸。推导的氨基酸序列55-184aa和365-487aa符合EGFR基因所具有的L1和L2受体结构域,在203-344aa和503-823aa含有EGFR家族特征区域CR1和CR2半胱氨酸富集区,并同为跨膜糖蛋白,在结构上具有一致性。经BlastP与GenBank已知物种氨基酸序列进行同源性比对,仿刺参EGFR基因氨基酸序列与果蝇EGFR相似性为49%,同源性为34%,与斑马鱼EGFR相似性为47%,同源性为34%,与淡水椎实螺EGFR相似性为49%,同源性为31%。据此推断,仿刺参EGFR基因属于EGFR家族成员。利用Real-time PCR技术检测了该基因在仿刺参各组织中的表达,结果显示在肠、呼吸树、表皮、体腔细胞、纵肌中EGFR均有表达,且体腔细胞和表皮表达量较高。结果表明,该基因可能在仿刺参组织发育和再生过程中起到重要的调控作用。 相似文献
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三疣梭子蟹细胞Cdk7基因克隆及其在卵巢发育中的表达 总被引：1，自引：1，他引：0
本实验应用RACE技术克隆获得了三疣梭子蟹(Portunus trituberculatus)周期蛋白依赖性激酶7(Cdk7)基因c DNA序列全长。基因5′和3′非编码区域(UTR)以及开放阅读框的长度分别是23 bp、178 bp和1056 bp,预测编码一个含有351个氨基酸,分子量为39.91 k D的蛋白质,包含一个丝氨酸/苏氨酸催化保守结构域和T-loop结构。同源性分析表明,三疣梭子蟹CDK7氨基酸序列与其他物种Cdk7相似度较高,说明Cdk7基因具有很好的保守性。实时荧光定量PCR结果显示,三疣梭子蟹Cdk7基因在卵巢组织中的表达量显著高于其他组织(P0.05)。Cdk7基因在卵巢发育不同时期的表达量存在差异,其在I期和Ⅱ期的表达量显著高于其他时期(P0.05)。去除眼柄后该基因在卵巢组织中的表达呈现先升高后降低的趋势,并于第4天达到最大值。本研究的结果表明,Cdk7基因参与了三疣梭子蟹卵巢发育调控,为深入开展三疣梭子蟹和其他甲壳动物性腺发育调控机理研究提供了重要信息。 相似文献
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采用RT-PCR和c DNA末端快速扩增技术(rapid amplification of c DNA ends,RACE)克隆出魁蚶过氧化氢酶(Sb CAT)基因c DNA全长序列,该基因全长为2181 bp,包括1431 bp的开放阅读框(open reading frame,ORF),96 bp的5'端非翻译区(UTR)和654 bp的3'-UTR。其中ORF编码477个氨基酸,预测分子量为54 ku,理论等电点为8.03。Sb CAT氨基酸序列与其他所选动物的氨基酸序列具有较高的相似性,其相似度为68%~96%,Sb CAT氨基酸具有CAT基因家族的特征性序列,包括CAT活性位点,1个亚铁血红素结合位点及3个催化位点残基。此外,Sb CAT还具有保守的亚铁血红素结合口袋与还原型辅酶Ⅱ(NADPH)结合位点。采用实时荧光定量PCR(q RT-PCR)检测了Sb CAT的组织表达特征。结果显示,Sb CAT m RNA在所检测的6种组织中均有表达,在外套膜中表达量较高,在肝胰腺和红细胞中表达量较低。经鳗弧菌和金黄色葡萄球菌刺激后,鳗弧菌刺激组的外套膜表达量一直较低,其他组织均表现出先上升再下降的趋势。研究表明,Sb CAT可能在魁蚶免疫防御中发挥重要作用。 相似文献
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在１５．０℃、１７．５℃和１９．０℃３个温度梯度下，各试验组纪体的生长速度和成活率６．３μｍ／ｄ，８．６μｍ／ｄ、９．３μｍ／ｄ；６１．０％、７０．５％、２３．０％。投附着基经２２ｄ培养各试验组稚贝规格（壳长×壳高）分别平均为６２０μｍ×５９５μｍ、７０５μｍ×６８０μｍ、７４５μｍ×７１０μｍ；各组生长速度和附着后成活率分别为１７．７μｍ／ｄ、２１．６μｍ／ｄ、２３．４μｍ／ｄ；３５．４％、３４ 相似文献
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Mrigal-common carp hybrids were produced and examined. Only few hybrids could be reared up to the age of 14 months. These did not show signs of maturity. The parents species mature in ponds in 6 months (common carp) or 12 months (mrigal). 相似文献
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Abstract – Similarities in the diets of larval, juvenile and adult black-stripe minnow ( Galaxiella nigrostriata ) and salamanderfish ( Lepidogalaxias salamandroides ), collected from ephemeral pools in south-western Australia during the mid-wet, late-wet and dry seasons, were assessed. During the mid-wet season, both intra- and interspecific competition for a diminishing food supply was minimised through differences in prey utilisation by the various developmental stages of both species. During the late-wet season, the pools contained only juvenile G. nigrostriata and juvenile and adult L. salamandroides , the diets of which were vastly different. A shift towards terrestrial fauna by G. nigrostriata and the ingestion of greater proportions of larger benthic prey taxa by L. salamandroides minimised interspecific competition for food at a time when densities of zooplanktors were rapidly declining. Timing differences in the onset of aestivation ensures that no interspecific competition for food occurs during the dry season. 相似文献
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Results are reported of the growth and yield of the Malawi chambo, Oreochromis karongae (Trewavas, 1941) association, and Oreochromis shiranus (Boulenger. 1896) in polyculture with Clarias gariepinus (Burchell. 1822). Six ponds of 150 m2 were assigned to two replicates of three treatments: C. gariepinus and O. shiranus; C. gariepinus and chambo spawned on station; C. gariepinus and chambo collected from the lake. Total fish stocking density was 3 m?2, and stocking ratio 1:2 (C. gariepinus: titapias). Initial mean body weights were 10.9 g for O. shiranus. 13.2 g for C. gariepinus and 13.3 g for chambo. The ponds were fertilized with chicken manure at the rate of 13.5 kg dry matter ha?1 day?1, representing the daily NPK loading of 0.618.0.186 and 0.191 kg ha?1, respectively. Maize bran was applied at 3% body weight day?1 for 168 days. Fish growth rate for C. gariepinus was significantly higher (P < 0.05) than that of the tilapias. Weight gains were 0.48-0.57 g day?1 for C. gariepinus, 0.30-0.31 g day?1 for chambo and 0.24 g day?1 for O shiranus. There was no difference (P > 0.05) in the growth of chambo despite the source of fingerlings. More fish biomass came from C. gariepinus stocked at 1 m?2 than from the tilapias stocked at 2 m?2. Extrapolated fish yields in the range of 4.37-4.75 t ha?1 year?1 were obtained from the polyculture of chambo and C. gariepinus. Oreochromis shiranus and C. gariepinus gave 3.95 t ha?1 year?1. Low water temperatures for 112 days slowed the growth of tilapias but did not seem to inhibit the growth of C. gariepinus. Chicken manure and maize bran, applied at smallholder farmer's level, has produced higher yields than by using O. shiranus in monoculture at optimal temperatures. The yields obtained show that the polyculture system is feasible during the cold season and may justify investment in aquaculture at semi-intensive level in Malawi. 相似文献
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Amusium laurenti is a functional hermaphroditewith well differentiated male and female gonadalareas. The gonadal index varies throughout the yearwith maximum values occurring in January, March andJuly, and minimum ones in August and September. Partial spawning (with respect to the individual andthe population) occurred from January to February andMarch to May. The major spawning event coincided withthe end of the period during which bottom watertemperature increased to temperatures above 26°C(July–August). Subsequently temperature decreased to22°C, with a period of gonadal restoration andincreasing gonadal index. Histological analysis ofthe gonads correlated with the gonadal index. 相似文献
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J. L. TOJO M. T. SANTAMARINA J. LEIRO F. M. UBEIRA M. L. SANMARTIN 《Journal of fish diseases》1994,17(2):135-143
Abstract. Various antiparasitic drugs belonging to several pharmacological groups were tested by bath administration for in vivo activity against a natural infestation of rainbow trout. Oncorhynchus mykiss (Walbaum), by he flagellate protozoan Ichthyobodo necator. Fish were also monitored for signs of drug toxicity. Complete elimination of infestation in all fish was achieved only by bithionol (25 mgl-1 for 3h on two consecutive days). Ketoconazole, toltrazuril, amprolium, sulphaquinoxaline, quinacrine, N-metylglucamine, chloroquine, 1,3-di-6-quinolylurea, diminazene aceturate and paromomycin were not effective. Diminazene aceturate (100 mgl-1 , 3h) was the only drug which was clearly toxic. The comparison of the response of symptomatic and asymptomatic infested fish to a formaldehyde bath indicates that poor health caused by I. necator may greatly increase the susceptibility of fish to the toxic effects of a drug or chemical. 相似文献
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Alfredo Salas‐Garza Griselda Parés‐Sierra Rodrigo Gómez‐Rigalt Eugenio Carpizo‐Ituarte 《Journal of the World Aquaculture Society》2009,40(4):460-471
This work presents the embryonic, larval, and postlarval development of Lithopoma undosa (L. undosa) (20 ± 2 C and the effect of natural inducers on the larval metamorphosis. Embryonic development until hatching of the trochophore larva took 15 h. Early veliger larvae with well‐developed shell were after 24 h. At 48 h, the operculum was completely formed. Cephalic tentacles were visible after 72 h. Larvae reached competence 6 to 7 d after fertilization, and was characterized by the presence of branched cephalic tentacles. The effect of natural inducers was evaluated. The percentage of metamorphosis with diatoms and adult mucus was 94% and 58% respectively, differing from the 25% without inducer. When was repeated, the metamorphosis was 100%, and 16% with benthic diatoms. The initial mean size of the postlarvae was 0.316 ± 0.02 mm. After 264 d the juveniles had attained a mean size of 5.98 ± 0.16 mm. These results will not only be of use in the evaluation of the culture of L. undosa, but also in studies on the characterization of inducers. 相似文献
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Amanda Pomposini Jonathan Blubaugh Robert Chad Boyce David T. Gauthier 《Journal of fish diseases》2019,42(5):739-749
Back Bay is an oligohaline, coastal bay in southeast Virginia, USA. Since 2004, leeches have been observed in the oral cavities of largemouth bass (Micropterus salmoides) in this body of water. Leeches (Myzobdella lugubris) have previously been documented in the oral cavities of largemouth bass in the Currituck Sound, which is confluent with Back Bay on its southern border. Supplemental stocking of largemouth bass in Back Bay since 2009 has resulted in an increasing population; however, concern exists that leech infestation may be negatively affecting health of larger fish, which are still less abundant than expected. Despite the wide distribution of this leech, there is little available literature regarding its health impacts on hosts. In this study, we examine potential impacts of oral leech infestations on stress markers and haematological parameters of largemouth bass in Back Bay. No significant changes in plasma glucose or cortisol were observed between leech‐infested and uninfested fish, and haematological parameters were not significantly different between the groups. Further, there was no evidence of systemic infections associated with leech infestation. 相似文献