共查询到19条相似文献，搜索用时 156 毫秒
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依据鸡白细胞介素2（chIL-2）全基因组序列设计引物探针,以简易基因组抽提法获得的鸭外周血单核细胞基因组DNA为模板,用长距离聚合酶链式反应（LD-PCR）扩增出鸭白细胞介素2（duIL-2）全基因片段。经序列测定,得到的duIL-2基因组DNA序列全长为3528bp。运用生物信息学方法对duIL-2的基因、启动子结构和cDNA序列编码的成熟蛋白进行系统分析,结果表明,duIL-2基因具有典型的4个外显子和3个内含子结构,和已知禽类及哺乳类同系物基因总体结构具有明显的相似性。鸭和鸡及其他哺乳动物IL-2基因的启动子序列具有显著的保守性,都具有AP-1、NF-AT、CD28RE、OCT、TATAbox转录起始因子结合位点和预测的转录起始位点。duIL-2cDNA序列编码的成熟蛋白进化分析表明,鸭和鹅的亲缘关系最近,和哺乳动物的亲缘关系较远,显示出明显的种属差异。 相似文献
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作者旨在克隆鸭GHSR基因mRNA部分编码区序列,并筛查克隆序列中的变异位点。采用RT-PCR法从巢湖鸭下丘脑组织中分离家鸭GHSR基因mRNA中编码区核酸序列,并选用30个个体cDNA,通过构建cDNA池对克隆编码区的序列变异测序检测。结果表明,克隆鸭GHSR基因mRNA部分编码区核酸序列长635 bp(GenBank登录号:EU005225),编码211个氨基酸,与鸡GHSR基因同源核酸相似性达到94%,氨基酸相似性为97%;cDNA池测序检测揭示克隆区段存在3个碱基变异位点,均为同义突变,未使编码氨基酸发生改变。克隆鸭GHSR基因mRNA编码区核酸、氨基酸序列与鸡同源序列的相似性,以及克隆核酸序列的变异检测结果表明,鸭GHSR基因在序列和功能上具有很高的保守性。 相似文献
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为研究胆绿素还原酶A(Biliverdin reductase A,BLVRA)基因的遗传分化,探索其结构和功能,根据鸡、人、小鼠、牛等动物BLVRA基因编码区的保守序列设计一对引物,以缙云麻鸭输卵管子宫部的总RNA为模板,利用RT-PCR技术扩增出鸭BLVRA基因的一段cDNA编码序列,并对其进行测序及序列分析。结果表明:该cDNA序列由634个核苷酸组成,编码211个氨基酸,分子量为24.1ku。与鸡、小鼠、牛、蟾蜍和人的核苷酸相似性分别为92.6%、64.9%、62.8%、69.0%、63.6%;氨基酸的相似性分别为96.2%、59.7%、60.7%、68.4%、59.7%,说明BLVRA基因在进化过程中较为保守。进一步的系统进化分析表明,鸭BLVRA基因与鸡的进化关系最近,蟾蜍次之,小鼠、牛和人较远,与传统的分类地位基本吻合。该基因部分cDNA序列的克隆,为获得BLVRA基因全长及其在各组织中的表达研究奠定了基础。 相似文献
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《畜牧与兽医》2016,(7):21-27
TGFBR1是TGF-β/Smad信号通路中的重要成员,在哺乳动物卵泡发育和排卵过程中发挥重要作用。本文通过克隆测序技术分离猪TGFBR1基因编码区序列,采用生物信息学方法对猪TGFBR1序列信息及其与哺乳动物其他物种间的进化和系统发育关系进行分析。结果发现:猪TGFBR1基因编码区序列全长1 512 bp,编码蛋白含有503个氨基酸残基,与哺乳动物其他物种的一致性分别在90%和95%以上;猪TGFBR1蛋白具有跨膜结构、GS区和激酶结构等保守结构域。进化分析显示哺乳动物TGFBR1基因的突变已达饱和状态,在进化过程中受到负选择的影响。基于TGFBR1基因编码区序列的系统发育分析发现,猪与同属偶蹄目的普通牛、绵羊的亲缘关系较近。 相似文献
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《动物营养学报》2015,(9)
本试验采用反转录聚合酶链式反应(RT-PCR)和cDNA末端快速扩增(RACE)技术克隆花鲈(Lateolabrax japonicus)调控摄食因子雷帕霉素靶蛋白(mTOR)、核糖体蛋白S6激酶beta-1(S6K1)、神经肽Y前体(NPY precursor)和脑肠肽前体(preproghrelin)cDNA全长序列。结果显示:花鲈mTOR(GenBank登录号KJ746670)cDNA全长8 296bp,开放阅读框7 551bp,编码2 516个氨基酸,预测其分子质量为286.14ku,与其他物种的相似性为62.0%~98.0%;S6 K1(GenBank登录号KJ746671)cDNA全长2 232bp,开放阅读框1 539bp,编码512个氨基酸,预测其分子质量为56.87ku,与其他物种的相似性为80.4%~84.9%;NPY precursor(GenBank登录号KJ850326)cDNA全长676bp,开放阅读框300bp,编码99个氨基酸,预测其分子质量为11.26ku,与其他物种的相似性为53.6%~99.0%;preproghrelin(GenBank登录号KJ850327)cDNA全长1 201bp,开放阅读框324bp,编码107个氨基酸,预测其分子质量为12.03ku,与其他物种的相似性为19.6%~85.0%。花鲈摄食调控因子cDNA全长序列地获得将为进一步研究其摄食调控机理奠定基础。 相似文献
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通过RT-PCR和RACE方法克隆了山羊Hspb10基因cDNA序列并进行了序列分析。结果表明:山羊Hspb10 cDNA全长1 056 bp,编码区全长780 bp,共编码259个氨基酸,提交至GenBank数据库中,收录号为JX067553。用Clustal W方法比对不同物种氨基酸序列同源性,山羊Hspb10与牛的同源性最高,核苷酸和氨基酸序列相似性分别为93.4%和96.5%;编码氨基酸序列BLAST比对结果也表明,哺乳动物Hspb10氨基酸序列极为保守,与禽类差异较大。获得山羊Hspb10 cDNA序列,可以为分子水平上研究山羊Hspb10的生物学功能奠定基础。 相似文献
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哺乳动物leptin的基因序列已被确认并成功克隆.尽管禽类“leptin cDNA”序列还未得到一致承认,但鸡leptin受体基因确实存在并已成功克隆,从另一方面证明了在禽类体内确实存在着与哺乳动物leptin结构和功能相似的分子.本文综述了禽类“leptin”发现及存在的争议,leptin受体结构以及哺乳动物leptin对禽类尿囊膜血管生成的影响. 相似文献
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试验旨在克隆天柱番鸭神经肽Y(neuropeptide Y,NPY)基因,并分析其在天柱番鸭不同组织中的转录水平。以天柱番鸭产蛋期组织混合cDNA为模板,通过RT-PCR扩增天柱番鸭NPY基因的完整CDS区进行克隆测序,并结合生物信息学分析工具分析其同源性及遗传进化关系,同时进行NPY蛋白理化特性、亚细胞定位、信号肽、糖基化与磷酸化位点、二级结构、三级结构等预测,并对NPY基因在天柱番鸭不同组织中的转录水平进行检测。结果表明,天柱番鸭NPY基因CDS区全长294 bp,编码97个氨基酸,核苷酸序列与氨基酸序列同源性比对显示,NPY基因在不同物种间具有一定的遗传多样性,与绿头鸭亲缘关系最近;NPY蛋白为酸性不稳定蛋白,存在1个信号肽(第1-28位氨基酸),亚细胞定位为100%位于细胞外;存在1个功能结构域PAH,同时含有2个O-糖基化和丰富的磷酸化位点,空间结构以α-螺旋和无规则卷曲为主。实时荧光定量PCR结果表明,NPY基因mRNA在天柱番鸭各组织中均有分布,在大脑中表达水平相对较高,在胰腺、腺胃中表达量次之,与其他组织间差异极显著(P<0.01),在肌胃中表达量最低。本试验结果为进一步研究NPY基因在调控家禽能量平衡、生长发育、脂肪沉积、繁殖性能等多种生理功能提供了参考。 相似文献
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试验旨在克隆天柱番鸭神经肽Y(neuropeptide Y,NPY)基因,并分析其在天柱番鸭不同组织中的转录水平。以天柱番鸭产蛋期组织混合cDNA为模板,通过RT-PCR扩增天柱番鸭NPY基因的完整CDS区进行克隆测序,并结合生物信息学分析工具分析其同源性及遗传进化关系,同时进行NPY蛋白理化特性、亚细胞定位、信号肽、糖基化与磷酸化位点、二级结构、三级结构等预测,并对NPY基因在天柱番鸭不同组织中的转录水平进行检测。结果表明,天柱番鸭NPY基因CDS区全长294 bp,编码97个氨基酸,核苷酸序列与氨基酸序列同源性比对显示,NPY基因在不同物种间具有一定的遗传多样性,与绿头鸭亲缘关系最近;NPY蛋白为酸性不稳定蛋白,存在1个信号肽(第1-28位氨基酸),亚细胞定位为100%位于细胞外;存在1个功能结构域PAH,同时含有2个O-糖基化和丰富的磷酸化位点,空间结构以α-螺旋和无规则卷曲为主。实时荧光定量PCR结果表明,NPY基因mRNA在天柱番鸭各组织中均有分布,在大脑中表达水平相对较高,在胰腺、腺胃中表达量次之,与其他组织间差异极显著(P0.01),在肌胃中表达量最低。本试验结果为进一步研究NPY基因在调控家禽能量平衡、生长发育、脂肪沉积、繁殖性能等多种生理功能提供了参考。 相似文献
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NPY、TSH-β、CaBP-28K基因在鸭繁殖期组织中的表达水平分析 总被引：1，自引：0，他引：1
本研究对繁殖期高邮鸭神经肽Y(NPY)、促甲状腺激素β(TSH-β)、钙结合蛋白-D28K(CaBP-28K)基因在中枢神经组织和周围组织表达量进行了实时荧光定量分析。结果表明:NPY基因在下丘脑中表达量显著高于在垂体和在小肠中的表达量(P<0.05),在卵巢、心脏、肝脏、脾脏、肾脏、胰腺、子宫、胸肌、腿肌等9个部位呈痕量表达;TSH-β基因在垂体、小肠中表达量较高,显著高于其他组织(P<0.05);CaBP-28K基因在小肠表达量最多,其次是卵巢和肾脏,但均显著大于垂体中表达量(P<0.05),在其他测定部位呈痕量表达,就生殖轴而言,CaBP-28KmRNA的表达量表现为卵巢>垂体>下丘脑。本研究为了解高邮鸭繁殖期相关基因的表达特征和内分泌机制,指导高邮鸭选种选育提供了理论依据。 相似文献
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禽流感病毒N3亚型神经氨酸酶基因的克隆 总被引：2，自引：0，他引：2
本研究应用11日龄SPF鸡胚增殖禽流感病毒标准株A/Duck/Germany/1215(H2N3),用LSTRIAZOLL试剂盒提取病毒RNA,应用自行设计的NA基因半特异性引物进行RT-PCR扩增,得到NA基因的全长DNA片段,克隆到PMD18-T载体上,并进行鉴定和序列测定.测定结果表明所获得的NA基因DNA片段长1451bp,编码469个氨基酸残基,与其他亚型的NA基因编码的氨基酸长度相一致.根据推导的氨基酸序列进行预测,该蛋白具有6个潜在的糖基化位点和19个半胱氨酸残基.与其他亚型的NA基因相比较发现克隆的NA基因符合神经氨酸的分子结构特征,本研究首次获得N3亚型NA基因全序列. 相似文献
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The objective of this study was to clone the sequence of duck type Ⅱ gonadotropin releasing hormone receptor (GnRHR-Ⅱ) gene, and to analyze the association of its expression levels with reproduction trait heterosis. RT-PCR method was used to clone GnRHR-Ⅱ gene fragments, and the expression levels during early laying days and laying fastigium period in Gaoyou duck, Jinding duck and crossed populations were tested by Real-time PCR method. Sequencing and homology analysis showed that the cloned duck GnRHR cDNA was about 500 bp in length, and it belonged to the avian type Ⅱ GnRHR isoforms. It was 96% identical to the cGnRHR-Ⅱ sequence reported in domestic chicken. From the early laying days to laying fastigium period, expression levels of GnRHR-Ⅱ gene in pituitary increased for Jinding duck, Gaoyou duck and crossed populations, with the highest expression levels in Jinding×Gaoyou hybrid group (P<0.01) . The expression levels of GnRHR-Ⅱ gene in hypothalamus also increased for Jinding duck, Gaoyou duck, however, it decreased in two crossed populations. The expression levels of GnRHR-Ⅱ gene in ovary were significantly higher for Jinding duck and two crossed populations than Gaoyou duck (P<0.01), but it decreased during laying fastigium period. Compared to Jinding duck and Gaoyou duck breeds, the two crossed populations had positive heterosis in 42-week production and 42-week egg fertilization percent, and negative heterosis in the first laying age and 42-week fertilized egg hatching rate traits. It suggested that the short-lived higher expression of ovary GnRHR-Ⅱ gene before early laying days be correlated with heterosis of the first laying age and 42-week production traits, and its higher expression levels in pituitary could aid to keep laying fastigium. 相似文献
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Although cDNA microarray studies have examined gene expression in human and rodent adipose tissue, only one microarray study of adipose tissue from growing pigs has been reported. Total RNA was collected at slaughter from outer subcutaneous adipose tissue (OSQ) and middle subcutaneous adipose tissue (MSQ) from gilts at 90, 150, and 210 d (n=5 age(-1)). Dye labeled cDNA probes were hybridized to custom porcine microarrays (70-mer oligonucleotides). Gene expression of insulin-like growth factor binding proteins (IGFBPs), hormones, growth factors, neuropeptide Y (NPY) receptors (NPYRs) and other receptors in OSQ and MSQ changed little with age in growing pigs. Distinct patterns of relative gene expression were evident within NPYR and IGFBP family members in adipose tissue from growing pigs. Relative gene expression levels of NPY2R, NPY4R and angiopoietin 2 (ANG-2) distinguished OSQ and MSQ depots in growing pigs. We demonstrated, for the first time, the expression of IGFBP-7, IGFBP-5, NPY1R, NPY2R, NPY, connective tissue growth factor (CTGF), brain-derived neurotrophic factor (BDNF) and ciliary neurotrophic factor (CNTF) genes in pig adipose tissue with microarray and RT-PCR assays. Furthermore, adipose tissue CTGF gene expression was upregulated while NPY and NPY2R gene expression were significantly down regulated by age. These studies demonstrate that expression of neuropeptides and neurotrophic factors in pig adipose tissue may be involved in regulation of leptin secretion. Many other regulatory factors were not influenced by age in growing pigs but may be influenced by location or depot. 相似文献
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K. Feng G.‐R. Zhang K.‐J. Wei G.‐W. Zou W.‐M. Wang 《Journal of animal physiology and animal nutrition》2014,98(2):338-346
Ghrelin, neuropeptide Y (NPY) and cholecystokinin (CCK) all have important roles in the regulation of feeding in fish and mammals. To better understand the role of the three peptides in appetite regulation in the early developmental stages of blunt snout bream (Megalobrama amblycephala), partial cDNA sequences of ghrelin, NPY and CCK genes were cloned. And then, real‐time quantitative PCR and RT‐PCR were used to detect and quantify the mRNA expressions of these genes from zygotes to larvae of 50 days after hatching (DAH). Ghrelin, NPY and CCK were all expressed throughout the embryonic and larval development stages, and the expression levels were higher in larval stages than in embryonic stages. Ghrelin and NPY mRNA expressions were upregulated at 1, 3, 5 DAH, while CCK mRNA expression was reduced significantly at 3 DAH. The mRNA expression levels of three genes in larvae varied significantly until 30 DAH. In adult fish, all three peptides were detected to be expressed in brain and several peripheral tissues. Ghrelin mRNA was mainly expressed in the intestine, whereas NPY and CCK mRNAs were mainly expressed in the brain. Taken together, these results indicate that ghrelin, NPY and CCK may have roles in early development and participate in the regulation of feeding of larvae in blunt snout bream and will be helpful for further investigation into feed intake regulation in adults of this species. 相似文献
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Shahein YE 《Veterinary immunology and immunopathology》2008,121(3-4):281-289
A full-length cDNA of an immunogenic protein was cloned from a cDNA library of the local Egyptian cattle tick Boophilus annulatus. Antibodies raised against B. annulatus larval proteins were used to screen a cDNA expression library. A 936bp cloned fragment was sequenced and showed an open reading frame of 516bp encoding a protein of 171 amino acids. Comparison of the deduced amino acid sequence with protein data bank revealed that the sequence is related to a sequence isolated from the hard tick Haemaphysalis qinghaiensis (Hq05). Southern blot analysis of B. annulatus genomic DNA showed that the cloned cDNA hybridized to double bands per restriction digest, suggesting that the cloned cDNA is a double copy gene. Amino acid analysis of the cloned gene revealed the presence of two casein kinase II phosphorylation sites in the N-terminal domain suggesting that this molecule may be involved in the signal transduction or gene expression pathways. RT-PCR and northern blotting revealed the presence of two isoforms of the Ba05 gene in salivary glands and in the 3-day-old eggs. The cloned gene without the signal peptide, was expressed in Escherichia coli under T7 promotor of pET-30b vector, and purified under denaturation conditions. The purified protein appeared as a single band on 12% SDS-PAGE with a molecular weight around 22.8kDa including the histidine tag of the vector. Antibodies raised against the purified molecule were used to detect the B. annulatus homologue to the Hq05 gene in whole tick, larvae and gut protein extracts. Immunoblotting revealed the presence of this molecule Ba05 only in whole tick and larval protein extracts and not in the gut protein extract. Using the same antibodies, homologues to the Ba05 gene were detected in other tick species as Hyalomma dromedarii and Rhipicephalus sp. but not in Ornithodoros moubata. 相似文献
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为建立鸭瘟病毒（DPV）快速、敏感的检测方法,本研究利用PCR技术扩增出DPV UL30基因中510bp的保守序列,并克隆到p MD18-T载体中作为标准品制作标准曲线,建立了DPV的SYBR Green I荧光定量PCR检测方法。该方法检测灵敏度可达5×101拷贝,与鸭细小病毒、鸭圆环病毒、小鹅瘟病毒、鸭肝炎病毒、鸭H9亚型流感病毒和鸭副粘病毒均不发生交叉反应,具有良好的特异性和重复性。结果表明,建立的实时荧光定量PCR具有特异、敏感、快速、定量、重复性好等优点,可用于临床DPV的检测。 相似文献
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为明确福建地区是否存在鹅出血性多瘤病毒（goose hemorrhagic polyomavirus,GHPV）感染,本研究对2016年福建省临床送检测19份鸭组织样品进行GHPV检测,结果从1例樱桃谷鸭中检测到GHPV感染阳性（记为GHPV-FJ201601株）。随后根据GenBank中GHPV参考株序列特征,设计针对GHPV的VP3基因特异性引物,利用PCR技术扩增获得GHPV的VP3基因片段。结果显示,GHPV-FJ201601株的VP3基因全长为654 bp,编码217个氨基酸。对编码的VP3蛋白分析发现其理论等电点为9.37,带负电荷氨基酸为23个（Asp+Glu）,带正电荷氨基酸为28个（Arg+Lys）;不稳定指数为38.43,是一个稳定蛋白;脂肪系数64.33,总平均疏水性指数为－0.744;该蛋白无典型的信号肽切割位点;其亚细胞定位类型为核定位;对其进行磷酸化分析发现,该蛋白存在14个氨基酸磷酸化位点。核苷酸同源性分析显示,不同来源GHPV代表株VP3基因相互之间核苷酸同源性均较高,均不低于99.7%。VP3基因遗传进化结果显示,GHPV-FJ201601株和鹅源GHPV分离株（匈牙利14234株与法国Toulouse Goose 2000株）处于同一遗传进化分支,与鸭源分离株（法国Toulouse Muscovy Duck 2008株、法国Toulouse Mule Duck 2008和中国106株）却处于不同的遗传进化分支,但所有GHPV分离株VP3基因遗传进化均较近。本研究首次证实福建地区樱桃谷鸭群中存在GHPV感染,为丰富不同地区与宿主的GHPV分子流行病学数据提供参考。 相似文献