共查询到17条相似文献，搜索用时 109 毫秒
1.
GH-IGF-I轴是鱼体体内一个重要的内分泌生理轴,主要调控鱼体的生长发育。用RT-PCR方法从加州鲈脑垂体和肝脏组织中分别扩增出加州鲈GH和IGF-I cDNA,克隆到pMD19 T-Vector上进行序列测定和分析。结果表明:1)加州鲈GH cDNA开放阅读框长为615 bp,编码204个氨基酸,其中信号肽17个氨基酸,成熟肽187个氨基酸。成熟肽中有四个保守的半胱氨酸残基(分别位于69,177,193,202),可形成两对二硫键。加州鲈GH氨基酸序列与蓝太阳鱼、斜带石斑鱼、金头鲷、虹鳟、鲤鱼、斑马鱼相比较,同源性分别为100%、97%、94%、66%、56%、53%。2)加州鲈IGF-I cDNA开放阅读框长为561bp,编码包括信号肽和B、C、A、D、E五个区域的186个氨基酸,形成成熟肽时,信号肽和E区域被切除。成熟蛋白的氨基酸序列与GenBank中已知的河鲈、三角鲂、金头鲷、舌齿鲈、斑马鱼的相比较,结果发现四个区域的保守性有差异,A区和B区保守性较高,C区和D区保守性较差。加州鲈GH和IGF-I cDNA的获得为进一步研究鱼体GH-IGF-I轴对生长发育的调控机制奠定了基础。 相似文献
2.
《广东海洋大学学报》2020,(3)
【目的】了解大刺鳅(Mastacembelus armatus)肌肉生长抑制素(myostatin)基因mstn的序列及其在生长发育中的调控功能。【方法】利用RACE方法克隆大刺鳅mstn全长cDNA序列,采用荧光定量PCR分析其在不同组织和胚胎发育阶段的表达情况。【结果与结论】大刺鳅mstn cDNA序列全长2 690 bp,包含200 bp 5′非编码区、1 359 bp 3′非编码区和1 131 bp开放阅读框,编码376个氨基酸。前22个氨基酸残基为信号肽,第37～254位氨基酸残基为TGF-β前肽域,第282～376位是TGF-β结构域,具蛋白酶水解位点RARR和C端生物活性区的9个保守半胱氨酸残基。系统发育分析表明,大刺鳅MSTN氨基酸序列与合鳃目、鲈形目鱼类同源性较高,与哺乳动物、鸟类、爬行类、两栖类同源性较低。实时定量PCR分析表明,大刺鳅mstn基因在各组织中均有表达,肌肉中表达量最高,眼、脑次之,鳃表达量最低;大刺鳅胚胎发育的各阶段mstn基因均有表达,囊胚期表达量最高,其次为多细胞期,出膜期表达量最低。 相似文献
3.
《广东海洋大学学报》2020,(5)
【目的】克隆大珠母贝(Pinctada maxima)胰岛素相关肽受体PmIRR基因,并分析其在不同组织中的表达。【方法】利用c DNA快速末端克隆技术(RACE)获得大珠母贝PmIRR基因cDNA全长序列。荧光定量PCR技术分析PmIRR在闭壳肌、鳃、外套膜边缘区、套膜区、中央区、足和肝胰腺等组织的表达模式。【结果与结论】PmIRR基因全长6 009 bp,5′非编码区(UTR)615 bp、3′UTR 764 bp、开放阅读框(ORF)长4 629 bp、编码1 542个氨基酸。序列分析表明,PmIRR含有保守结构域Fu、3个FNⅢ结构域和Tyrkc结构域,并含有信号肽和2个跨膜结构域。PmIRR在大珠母贝各个组织存在差异表达,在鳃、肝胰腺和外套膜边缘区中显著高表达。 相似文献
4.
用RT-PCR和RACE方法,从荷那龙罗非鱼(Oreochromis hornorum)垂体中克隆到生长激素促分泌素受体(GHSR)cDNA全序列。荷那龙罗非鱼GHSR基因具有GHSR-1a与GHSR-1b两个高度保守cDNA序列。GHSR-1a序列全长1 646 bp,包括225 bp的5′非编码区,266 bp的3′非编码区和1 155 bp的开放阅读框,编码384个氨基酸残基,具有7个跨膜结构域结构(transmembrane domains,TM);GHSR-1b序列全长1 877 bp,包括225 bp的5′非编码区,755 bp的3′非编码区和897 bp的开放阅读框,编码298个氨基酸残基,只具有前5个TM,在第6个TM的第4个氨基酸处开始缺失。将GHSR cDNA序列与基因组序列比较发现,这两种cDNA转录本来自同一个基因的不同变体。 相似文献
5.
【目的】克隆马氏珠母贝(Pinctada martensii)法尼酸甲基转移酶(FAMe T)基因,并分析其在各组织中的表达。【方法】利用cDNA末端快速扩增技术(RACE)克隆获得马氏珠母贝FAMeT(PmFAMeT)基因的cDNA全长序列,利用实时荧光定量PCR(qPCR)方法分析PmFAMeT在马氏珠母贝不同组织中的表达模式。【结果与结论】PmFAMeT包含5′非编码区120 bp,3′非编码区968 bp和开放阅读框(ORF)1 536 bp,编码511个氨基酸。序列分析表明,PmFAMeT含有信号肽序列和跨膜结构域,并有WSC结构域、TSP1结构域和2个Methyltransf_FA结构域。将推导的PmFAMeT氨基酸序列与其他物种的FAMeT序列进行比对发现,不同物种的Methyltransf_FA序列同源性较高。PmFAMeT在马氏珠母贝的各个组织中均有表达,且在边缘膜、肝胰腺和性腺中的表达量显著高于其他组织。 相似文献
6.
【目的】克隆马氏珠母贝(Pinctada martensii)肿瘤坏死因子受体相关死亡域蛋白(TRADD)基因,并分析其在各组织中的表达。【方法】利用cDNA末端快速扩增技术(RACE)克隆获得马氏珠母贝PmTRADD基因的c DNA全长序列,利用实时荧光定量PCR(qPCR)方法分析PmTRADD基因在马氏珠母贝不同组织中的表达模式。【结果与结论】PmTRADD包含5′非编码区101 bp,3′非编码区144 bp和开放阅读框(ORF)591 bp,编码196个氨基酸。序列分析表明,PmTRADD没有信号肽和跨膜结构域,C端含有一个死亡结构域(DEATH)。将PmTRADD死亡结构域的氨基酸序列与其他物种的TRADD死亡结构域序列进行比对,发现不同物种的TRADD死亡结构域序列同源性较低。PmTRADD在马氏珠母贝各组织中均有不同程度表达,在鳃组织中表达最高,肝胰腺次之,闭壳肌中基本无表达。 相似文献
7.
五种鲤科鱼类生长激素cDNA的克隆和序列分析 总被引：1，自引：0，他引：1
通过RT-PCR方法,以草鱼、鳙鱼、鲫鱼、鲤鱼、齐口裂腹鱼等5种鲤科重要经济鱼类的垂体总RNA为模板扩增出其生长激素(fish growth hormone,fGH)的完整ORF序列,并克隆到pMD18-T载体上,命名为pMD-1(草鱼)、pMD-2(鳙鱼)、pMD-3(鲫鱼)、pMD-4(鲤鱼)、pMD-5(齐口裂腹鱼)。测序结果显示,ORF序列其长度均为633 bp。序列分析表明,所有ORF序列均以ATG为起始密码,以TAG为终止密码,编码210个氨基酸残基,推导的生长激素前体由22个氨基酸的信号肽和188个氨基酸的成熟肽组成。同源性分析表明,草鱼、鳙鱼、鲤鱼、鲫鱼和齐口裂腹鱼的fGH同源性在93.3%~99.5%之间。 相似文献
8.
【目的】探究马氏珠母贝接头蛋白CIKS(PmCIKS)在马氏珠母贝免疫反应中的作用。【方法】采用cDNA末端快速扩增(RACE)技术获得PmCIKS基因cDNA全长序列,运用生物信息学手段分析该序列,用实时荧光定量PCR(RT-PCR)技术检测PmCIKS基因在马氏珠母贝7个组织中的表达模式。【结果】PmCIKS基因cDNA全长为1 987 bp,其中5′UTR长为228 bp,3′UTR长为148 bp,包含24 bp的ploy A,开放阅读框(ORF)为1 611 bp,编码536个氨基酸,预测其分子质量约为61.3 ku,等电点为7.01。物种间CIKS有较高的保守性。PmCIKS在马氏珠母贝肝胰腺、性腺、闭壳肌、鳃、血细胞、外套膜和足中均有表达,其中在血细胞中表达量最高。 相似文献
9.
《广东海洋大学学报》2017,(3)
以RACE技术克隆获得马氏珠母贝载脂蛋白-2(Pm-ApoL2)基因cDNA全长序列,以传统萃取法提取黄色与白色闭壳肌个体的类胡萝卜素,以荧光定量技术检测Pm-ApoL2基因在各个组织中的表达量及黄与白闭壳肌个体的Pm-ApoL2表达量。结果表明,Pm-ApoL2基因cDNA序列全长1 328 bp,开放阅读框(ORF)长705 bp,编码234个氨基酸,5′非翻译区(5′UTR)长342 bp,3′UTR长281 bp。Pm-ApoL2在肝胰腺中表达量最高,其后依次是外套膜、鳃、闭壳肌,其中肝胰脏表达量显著大于其它3个组织(P0.05)。黄、白色闭壳肌个体之间的类胡萝卜素含量和Pm-ApoL2的表达量都存在显著性差异(P0.05),且二者的类胡萝卜素含量与Pm-ApoL2表达量呈显著正相关。Pm-ApoL2基因为马氏珠母贝类胡萝卜素代谢候选基因。 相似文献
10.
《广东海洋大学学报》2017,(4)
Lec-8(C-type lectin 8)属于C-型凝集素超家族,在机体抵抗微生物的感染过程起重要的作用。本研究采用RACE技术克隆出了PmLec-8基因cDNA全长序列,并且运用实时荧光定量PCR(qRT-PCR)技术检测了PmLec-8基因在马氏珠母贝(Pinctada fucata martensii)不同组织中的表达模式。结果显示:PmLec-8基因序列全长737 bp,其中5′UTR长为36 bp,3′UTR长为113 bp,开放阅读框(ORF)为588 bp,编码195个氨基酸,预测其分子质量约为22.90 ku,等电点为5.17;PmLec-8具在一个糖类识别结构域(Carbonhydrate-recognition domain,CRD),符合典型的C型凝集素(C-type lectin)家族特征;多序列比对结果表明物种间Lec-8的同源性较低,4个半胱氨基酸残基高度保守;qRT-PCR数据分析表明,PmLec-8在马氏珠母贝肝胰腺、性腺、闭壳肌、鳃、血细胞和外套膜中均有表达,其中肝胰腺中表达量最高,其次是鳃,差异具统计学意义(P0.05)。 相似文献
11.
Lectins play a crucial role in the innate immunity of invertebrates and vertebrates by recognizing and disposing of pathogens.We obtained the complete cDNA of a C-type lectin(EALec1) from Epinephelus akaara using RACE.The complete EALec1 cDNA sequence was 827 bp.The 5-UTR and 3-UTR were 28 bp and 151 bp,respectively,in length.The sequence also contained a polyadenylation signal AATAAA and a poly(A) tail.The EALec1 cDNA encodes polypeptides with 215 amino acids,including a signal peptide of 31 amino acids.Th... 相似文献
12.
将大口黑鲈(Micropterus salmoide)肌肉生长抑制素(Myostatin,MSTN)前肽(MSTN-Pro)的cDNA定向克隆到真核表达载体pcDNA3.1(-)/mycHisB中,双酶切检测和测序鉴定证实,插入pcDNA3.1(-)/mycHisB载体中的片段为目的基因的核苷酸序列,MSTN基因前肽cDNA为正向插入,且重组质粒无错配或插入移位等突变。采用肌肉注射法将重组表达质粒注入大口黑鲈背部肌肉组织,在注射后第2天经RT-PCR检测到MSTN前肽基因mRNA的表达,第6天经免疫组化学检测到MSTN前肽蛋白的表达,第8天蛋白表达强度增强,对照组始终未检测到MSTN前肽基因的表达。 相似文献
13.
以勒氏笛鲷(Lutjanus russellii)基因组DNA为模板,采用同源克隆的方法,获得2 887 bp的肌肉生长抑制素(myostatin,MSTN)基因组序列,该MSTN序列具有3个外显子和2个内含子,包括101 bp的5′-UTR、385bp的外显子1、354 bp的内含子1、370 bp的外显子2、761 bp的内含子2、381 bp的外显子3和1 932 bp的3′-UTR。整个开放阅读框编码了378个氨基酸,前面的22个氨基酸为信号肽,具有9个保守的半胱氨基酸及一个RVRR的蛋白酶解加工位点。氨基酸序列分析发现,该基因编码的蛋白质与其他鱼类的I型同源性较高,与其他鱼类的2型MSTN同源性较低,且与鲈形目的同源性最高,与鲤形目的同源性较低,与人、鼠和鸡的同源性最低。采用邻接法(Neighbor-Joining)构建的MSTN的系统发育树表明,勒氏笛鲷MSTN与鲈形目鱼类的狼鲈属亲缘关系较近,且与鱼类的MSTN-1聚为1支。这表明该基因属于Ⅰ型MSTN基因。 相似文献
14.
AMP deaminase catalyzes the conversion of AMP into IMP and ammonia. In the present study, a full-length cDNA of AMPD1 from skeletal muscle of Japanese flounder Paralichthys olivaceus was cloned and characterized. The 2 526 bp cDNA contains a 5’-UTR of 78 bp, a 3’-UTR of 237 bp and an open reading frame (ORF) of 2 211 bp, which encodes a protein of 736 amino acids. The predicted protein contains a highly conserved AMP deaminase motif (SLSTDDP) and an ATP-binding site sequence (EPLMEEYAIAAQVFK). Phylogenetic analysis showed that the AMPD1 and AMPD3 genes originate from the same branch, but are evolutionarily distant from the AMPD2 gene. RT-PCR showed that the flounder AMPD1 gene was expressed only in skeletal muscle. QRT-PCR analysis revealed a statistically significant 2.54 fold higher level of AMPD1 mRNA in adult muscle (750±40 g) compared with juvenile muscle (7.5±2 g) (P<0.05). HPLC analysis showed that the IMP content in adult muscle (3.35±0.21 mg/g) was also statistically significantly higher than in juvenile muscle (1.08±0.04 mg/g) (P<0.05). There is a direct relationship between the AMPD1 gene expression level and IMP content in the skeletal muscle of juvenile and adult flounders. These results may provide useful information for quality improvement and molecular breeding of aquatic animals. 相似文献
15.
Shuo Li Chunmei Li Xubo Wang Yanan Wang Zhipeng Liu Teng Zhai Quanqi Zhang 《中国海洋大学学报(英文版)》2010,9(1):59-64
A novel immune-related gene was expressed in Japanese flounder (Paralichthys olivaceus) injected with Vibrio anguillarum. The complete cDNA contained a 169 bp 5’UTR, a 336 bp open reading frame (ORF) encoding 111 amino acids and a 556bp 3’UTR.
Six exons and five introns were identified in the PoIR2 gene. Blastp similarity comparison showed its encoding protein had 50% similarity to Danio rerio neuromedin S (NMS), but further alignment indicated they did not have NMS C-terminal conservational signature domain. So
it was not defined as an NMS homologue. Protein structure analysis indicated it had a 26aa signal peptide and was a secretory
pathway protein. RT-PCR demonstrated that the expression of PoIR2 was quickly induced and drastically increased in liver, kidney, spleen, gills, intestine, heart, and skeletal muscle after
infected with V. anguillarum. These results indicated that the PoIR2 might play some important role in Japanese flounder immune response system. This gene was named PoIR2 (P.olivaceus immune-related gene 2, GenBank accession number: EU224372). The mature PoIR2 peptide was expressed in BL21(DE3) pLysS using pET-32a(+) vector and a great part of the recombinant mature peptide existed
as soluble type. 相似文献
16.
Acting as one of the pattern recognition receptors (PRRs), C-type lectin is believed to mediate pathogen recognition and plays an important role in the clearance of pathogens as part of the innate immune system. In this work, a novel C-type lectin gene (named LvLec1) was cloned from the shrimp Litopenaeus vannamei. The ORF of LvLec1 is 510 bp, encoding 169 amino acids. The deduced amino acid sequence contains a putative signal peptide of 19 amino acids at the N-terminal and a carbohydrate recognition domain... 相似文献
17.
《中国海洋大学学报(英文版)》2017,(5)
Lymphocystis disease virus(LCDV) infects target cells by attaching to a 27.8 k Da receptor(27.8R) protein in flounder Paralichthys olivaceus, and anti-27.8R monoclonal antibodies(MAbs) have been developed. However, the 27.8R existence in tissues of sea bass(Lateolabrax japonicus) and its role in LCDV infection have remained unclear. In this study, the results of western blotting demonstrated that the same 27.8R was shared by flounder and sea bass. LCDV-free sea bass individuals were intramuscularly injected with LCDV, and viral copies were detected in tissues from 3 h post infection and showed a time-dependent increase during 9 days infection. Distribution and synthesis of 27.8R in sea bass tissues were investigated by using anti-27.8R MAbs as probes. It was found that 27.8R was distributed in all the tested tissues. The levels of 27.8R protein were highest in gill and skin, then a bit lowly in stomach, head kidney and heart, followed by spleen, intestine, blood cells, gonad and liver, and least in kidney and brain in healthy sea bass. Upon LCDV infection, 27.8R synthesis was up-regulated in each tissue, and higher in the tissues with higher LCDV copies. The 27.8R and LCDV were detected in some peripheral blood leukocytes but not in red blood cells. These results suggested that 27.8R was widely distributed in sea bass tissues, and it served as a receptor and correlated with tissue tropism of LCDV infection. Furthermore, leukocytes had the potential of being a LCDV carrier and were responsible for a systemic infection of LCDV in sea bass. 相似文献