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Xia P Liu Y Liu X Zhang Z Duan E Lu X Zhao J Cui B 《Veterinary immunology and immunopathology》2011,141(1-2):144-150
Receptors for the Fc fragment of IgG (FcγRs) constitute one of the main effector mechanisms through which IgG immune complexes exert their action. Four FcγRs, FcγRI (CD64) with high affinity, FcγRI with intermediate affinity, FcγRII (CD32) and FcγRIII (CD16) with low affinity, have been identified. There are three FcγRII isoforms (activating FcγRIIa and FcγRIIc, and inhibiting FcγRIIb) existing in humans, one isoform in mice (inhibiting FcγRIIb), and two isoforms in cattle (inhibiting FcγRIIb, activating FcγRIIc). Two splice sub-isoforms of FcγRIIb, FcγRIIb1(b1) and FcγRIIb2(b2), have been identified in humans, mice and cattle, however, few of FcγRIIb sub-isoforms have been investigated in pig. In this study, we describe the molecular cloning, sequencing and characterization of a porcine FcγRIIb sub-isoform, FcγRIIb1. The cDNA encoding porcine FcγRIIb1 was isolated from peripheral blood leucocytes RNA with RT-PCR. The porcine FcγRIIb1 cDNA contains a 951bp open-reading frame, encoding a 316 amino acid transmembrane glycoprotein composed of two immunoglobulin (Ig)-like extracellular domains, a transmembrane region and a cytoplasmic tail with an immunoreceptor tyrosine-based inhibiting motif (ITIM). The porcine FcγRIIb1 shares 98.3% homology and has a 19 amino acid in-frame insertion in cytoplasmic tail when compared with amino acid sequence of DQ026064. Immunofluorescence analysis showed that the glycoprotein encoded by the porcine FcγRIIb1 cDNA was expressed in the stable transfected COS-7 cells, and an immunoglobulin-binding assay showed that it had binding activity for IgG immune complexes. Identification of the porcine FcγRIIb1 will help our understanding of the molecular basis of IgG-FcγR interaction in the porcine immune response. 相似文献
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克隆了猪NPM1基因的cDNA和第4内含子序列，并利用生物信息学方法进行验证。所得cDNA序列全长1195bp且包含一个完整的开放阅读框，编码294个氨基酸。内含子序列长300bp，遵循GT／AG剪接法则。序列分析表明，该基因与已报道的人、大鼠和小鼠等物种的NPM1基因高度同源，氨基酸序列同源性分别为95％、91％和91％。该基因编码的蛋白具有Nucleoplasmin保守功能域。用体细胞杂种板将该基因定位在猪16号染色体q21区段，辐射杂种板定位结果表明该基因与猪16号染色体上SW977标记紧密连锁。 相似文献
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Yunchao Liu Aiping Wang Songlin Qiao Gaiping Zhang Jun Xi Leiming You Xiaohui Tian Qiaomu Li Lina Zhang Junqing Guo 《Veterinary immunology and immunopathology》2010,133(2-4):243-249
Immunoglobulin G (IgG) Fc receptors (FcγRs) bind to immune complexes through interactions with the Fc region of IgG to initiate or inhibit the defense mechanism of the leukocytes on which they are expressed. In this study, we describe the cloning, sequencing and characterization of ovine FcγRII. By screening a translated expression sequence tag (EST) database with the protein sequence of bovine IgG Fc receptor II, we identified a putative ovine homologue. Using rapid amplification of cDNA ends (RACE), we isolated the cDNA encoding ovine FcγRII from peripheral blood leucocyte RNA. The ovine FcγRII cDNA contains an 894 bp open-reading frame, encoding a 297 amino acid transmembrane glycoprotein composed of two immunoglobulin-like extracellular domains, a transmembrane region and a cytoplasmic tail with an immunoreceptor tyrosine-based inhibitory motif (ITIM). The glycoprotein encoded by the cloned cDNA was then expressed on the surface of COS-7 cells and immunoglobulin-binding assays show that it binds ovine IgG1, but not IgG2. Identification of the ovine FcγRII will aid in the understanding of the molecular basis of IgG–FcγR interaction. 相似文献
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Cloning and characterization of porcine resistin gene 总被引：5，自引：0，他引：5
Dai MH Xia T Chen XD Gan L Feng SQ Qiu H Peng Y Yang ZQ 《Domestic animal endocrinology》2006,30(2):88-97
Resistin is a member of resistin-like molecules (RELMs) and a hormone secreted from mature adipocytes in rodents and leukocytes in human. We now report the cloning and characterization of the full-length porcine resistin cDNA and gene. Sequence analysis indicated that the pig resistin cDNA sequence had an open reading frame of 330 bp encoding a 12 kDa protein of 109 amino acids. The deduced amino acid sequence showed 75.2% identity to the human resistin. The porcine resistin gene was composed of four exons and had exactly the same exon structure as the human resistin gene. The tissue distribution of porcine resistin mRNA was assessed by semi-quantitative RT-PCR. Resistin gene expression was the highest in porcine leukocytes and low in adipose tissue. Resistin protein could be detected in porcine serum by western blotting and it circulated in serum as dimers and trimers. We provided the first evidence that resistin was abundantly expressed in porcine leukocytes and had an expression pattern similar to that in human resistin mRNA and protein. This suggests that the pig may be a suitable animal model for studying the function of resistin in human insulin resistance. 相似文献
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The receptor I for the Fc region of immunoglobulin G (Fc gamma RI) is a member of the Ig superfamily with a high affinity, and it mediates antibody-dependent cellular cytotoxicity and immune complex clearance. In this study, a cDNA encoding the bovine Fc gamma RI was cloned. The full-length cDNA sequence is 1050 bp long with a short 5'- and long 3'-untranslated end regions, which codes for 349 amino acids and contains a signal peptide, an extracellular region with three Ig-like domains, and transmembrane and intracytoplasmic domains. Five potential N-linked glycosylation sites are recognized in this sequence. Compared with the sequences of human and mouse Fc gamma RI, the homologies of nucleotide sequences are 80 and 69% and homologies of deduced amino acid sequences are 66 and 55%, respectively. It is shown that the sequences of the monomeric IgG binding domain in these three species of Fc gamma RI are highly conserved. 相似文献
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为研究猪肺巨噬细胞FcγR Ⅲ的生物学功能,本研究应用RT-PCR技术从猪肺巨噬细胞总RNA中克隆出猪FcγR Ⅲ的cDNA序列,并对其进行了分析。结果表明,克隆到的序列长820 bp,包含有1个771 bp完整开放阅读框(ORF),与GenBank中登录的猪FcγR Ⅲ序列(AF237453)的核苷酸同源性为99.9％;与人、牛、马、绵羊、猕猴、狗、猫、小鼠氨基酸同源性分别为61.6%、62.9%、55.3%、62.2%、63.0%、59.0%、61.8%和53.2%;蛋白质分子结构预测结果表明,该分子由信号肽（20个氨基酸）、胞外区(185个氨基酸)、跨膜区(23个氨基酸)和胞内区(28个氨基酸)组成,在胞外区存在2个Ig样结构域。猪肺巨噬细胞FcγR Ⅲ基因的成功克隆,为进一步研究其结构与功能奠定基础。 相似文献
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研究旨在对猪T细胞诱导型刺激物(ICOS)基因的cDNA序列进行克隆与分析。根据已报道的人ICOS基因cDNA序列设计引物,首次从猪脾脏组织总RNA中扩增出ICOS基因编码区全长cDNA序列,克隆于pMD18-T载体后进行测序并进行序列拼接,运用生物信息学分析DNA序列。结果表明:该基因编码区全长630bp,编码210个氨基酸,包含5个外显子。该序列与人全基因核苷酸序列及推导的氨基酸序列的同源性分别为80%和85%;与狗和小鼠的推导氨基酸序列的同源性分别为81%和75%。这为进一步研究该基因的结构特点和功能奠定了良好基础。 相似文献
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根据GenBank已收录的牛(Bos taurus)、人(Homo sapiens)和小鼠(Mus musculus)等物种Ets-1基因序列的同源保守区域,设计特异性引物,采用RT-PCR和RACE技术,分离并克隆了西农萨能奶山羊(Capra hircus)Ets-1基因的cDNA序列。该序列全长2 263 bp(GenBank登录号HQ589338),包括5’UTR 331 bp,CDS 1 326bp和3’UTR 606 bp,编码441个氨基酸组成的蛋白质。核苷酸序列分析发现,山羊编码序列与牛、猪、人、小鼠等的相应序列同源性分别为98%、94%、92%和90%,3’UTR相应序列为96%、83%、81%和77%,5’UTR相应序列为98%、85%、82%和71%。氨基酸序列分析发现,山羊与牛、猪、人和小鼠的Ets-1的相似性较高,均在95%以上。蛋白质结构分析发现,其蛋白质分子量为50 340.8 D,等电点为5.08,具有典型的螺旋-转角-螺旋结构域,不存在跨膜结构,并且整个序列不含信号肽。 相似文献
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Liang Tang Craig Sampson Matthew J. Dreitz Catherine McCall 《Veterinary immunology and immunopathology》2001,80(3-4):259-270
cDNAs encoding four different canine immunoglobulin G (caIgG) γ chains were identified in this study. One of these IgG γ chain cDNAs, (caIgG-A), represents 92.5% of the IgG γ chain cDNAs in a dog spleen cell cDNA library; a second partial IgG γ chain cDNA (caIgG-B) was also identified in the library. The other two IgG γ chain cDNAs (caIgG-C and caIgG-D) were RT-PCR amplified from canine lymphoma samples. Comparison of the four different canine IgG γ chain cDNAs showed homologies from 83.6 to 89.2% and from 73.1 to 81.8% at nucleotide and amino acid sequence levels, respectively. Despite the high similarity in CH1, CH2 and CH3 domains among the different caIgG γ chains, the hinge regions were distinct, sharing only 19.0–35.2% homology at the amino acid level. No multiple duplication of the hinge region, as reported for human IgG1 and IgG3, was detected in any of the canine IgG γ chains. The numbers of cysteines in the putative hinge regions were found to be 3, 2, 7 and 3 for the four canine IgG heavy γ chains (A, B, C and D), respectively. Specific primers were designed based on caIgG γ chain hinge region DNA sequences and were used in RT-PCR for measuring different caIgG γ chain mRNA levels in canine PBMC samples. 相似文献
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为了解猪Toll样受体5(TLR5)蛋白的结构特征和进化关系,本研究从肠系膜淋巴结组织总RNA中克隆出猪Toll样受体5基因的cDNA序列。序列全长2641bp,其中2571bp的开放阅读框编码856个氨基酸残基的猪TLR5,含17.4%的亮氨酸,并有一段19个氨基酸的信号肽序列;同源性分析结果显示,TLR5在进化过程中具有高度保守性,与人、牛、山羊、绵羊、小鼠和大鼠的氨基酸序列同源性分别为77.5%、79.6%、78.3%、81.0%、69.2%和68.9%。蛋白分子结构预测结果表明,该分子由胞外区(642个氨基酸)、跨膜区(23个氨基酸)和胞内区(191个氨基酸)组成,胞外区具有LRR结构域,胞内区具有TIR结构域,表现出典型的TLR家族结构特征。表明猪TLR5分子具有病原分子模式识别和信号传导的作用,这为其结构与功能的进一步研究奠定了基础。 相似文献
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Immunobiotic Lactobacillus strains augment NLRP3 expression in newborn and adult porcine gut-associated lymphoid tissues 总被引：1，自引：0，他引：1
Tohno M Shimosato T Aso H Kitazawa H 《Veterinary immunology and immunopathology》2011,144(3-4):410-416
We isolated cDNA encoding porcine nucleotide-binding domain-like receptor family, pryin domain containing 3 (NLRP3) from Peyer's patches. The complete nucleotide open reading frame of porcine NLRP3 contains 3108-bp encoding a deduced polypeptide of 1036-amino acid residues. The porcine NLRP3 amino acid sequence is more similar to the longest isoform of human than the mouse counterpart. The predicted amino acid sequence of porcine NLRP3 presented nine C-terminal leucine-rich repeat domains. In newborn swine, the expression of NLRP3 was detected at higher levels in spleen and mesenteric lymph nodes, while lower levels were observed in intestinal tissues. In adult swine, NLRP3 was strongly expressed in Peyer's patches and the mesenteric lymph nodes, and the expression level in the lower intestinal tissues was comparable to that in spleen. Toll-like receptor and nucleotide-binding domain ligands, as well as Lactobacillus delbrueckii subsp. bulgaricus and Lactobacillus gasseri, enhanced NLRP3 expression in gut-associated lymphoid tissues (GALT) of newborn and adult swine. Our results should aid in understanding the intestinal immunoregulatory mechanisms underlying NLRP3 activation and the priming ability of immunobiotic lactic acid bacteria in porcine GALT. 相似文献
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E Kim J‐S Kim Y Lee B‐S Song B‐W Sim S‐U Kim T Saitoh H Yazawa T Nunoya K‐T Chang 《Reproduction in domestic animals》2013,48(1):90-97
IZUMO1, belonging to the family of mammalian immunoglobulin proteins, has been well characterized in the mouse. Here, we describe the molecular cloning and expression analysis of porcine IZUMO1 (pIZUMO1). Partial sequence information published in the National Center for Biotechnology Information (NCBI) database was used to generate the full‐length sequence for IZUMO1 using rapid amplification of cDNA ends (RACE). A search of the porcine genomic sequence in the NCBI database identified a bacterial artificial chromosome (BAC) encoding the pIZUMO1 gene. This BAC is derived from porcine chromosome 6 and is syntenic with the corresponding regions of mouse, bovine, and human genomes encoding the IZUMO gene family. This BAC was found to encode an IZUMO1 protein with a predicted amino acid sequence having high similarity with mouse and human IZUMO1. Western blot analysis of proteins from porcine tissues indicated that pIZUMO1 was specifically expressed in the sperm. Furthermore, to confirm whether pIZUMO1 forms complexes, we overexpressed pIZUMO1 in HEK293 cells. The recombinant pIZUMO1 from cell extracts was found to form complexes. Our finding suggests that pIZUMO1 forms homodimeric complex on the sperm membrane. Furthermore, an IVF inhibition assay with an antibody for the porcine IZUMO1 Ig‐like domain showed that Ig‐like domain effectively prevented pig sperm–egg interactions. 相似文献
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为了进一步了解猪nectin-2基因的结构与功能,本研究采用生物信息学结合RT-PCR的方法从猪脑组织中克隆到了猪nectin-2基因,并对其核苷酸序列和推导的氨基酸序列进行了比较分析。猪nectin-2基因的编码区含有1440个核苷酸,编码479个氨基酸,其中信号肽由32个氨基酸组成,胞外域由330个氨基酸组成,含有2个潜在的N-糖基化位点和6个半胱氨酸残基,跨膜区由23个氨基酸组成,胞浆区由94个氨基酸组成,猪nectin-2基因与犬、马、家鼠、人、恒河猴、牛、黑猩猩的nectin-2基因核苷酸序列同源性分别为85.4%、85.7%、78.6%、82.1%、82.1%、81.9%和82.1%;推导氨基酸序列的同源性分别为84.5%、83.0%、74.7%、75.7%、76.4%、78.4%和75.5%。本试验为进一步深入研究猪伪狂犬病病毒与宿主之间的关系奠定了基础。 相似文献
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Sanz G Jiménez-Marín Á Barbancho M Garrido JJ 《Veterinary immunology and immunopathology》2012,145(1-2):505-510
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de la Lastra JM Shahein YE Garrido JJ Llanes D 《Veterinary immunology and immunopathology》2003,93(3-4):107-115
CD97 is a member of a novel subfamily of leukocyte proteins that are characterized by the presence of tandemly repeated extracellular epidermal growth factor (EGF)-like domains and a seven-span transmembrane region, known as EGF-TM7. We here report the cloning of cDNA encoding the pig homologue of CD97. A pig CD97 specific probe was generated by PCR amplification of pig leukocyte cDNA, using primers based on consensus regions among the known sequences of mouse and human CD97. Screening of a pig aorta smooth muscle cDNA library identified one clone containing an open reading frame (ORF) that encoded an 18 amino acid putative signal peptide, a 141 amino acid sequence consisting of three EGF domains, a mucin-like spacer region of 276 amino acid, containing a G-protein coupling motif of 52 amino acids, followed by a 250 amino acid region containing seven membrane spanning domains and a 47 amino acid cytoplasmic tail. The amino acid sequence of the clone was 75, 67 and 59% homologous to cattle, human and mouse CD97 antigen, respectively. Therefore, it was termed pig CD97. Pig CD97 antigen shares many structural features with human, cattle and mouse CD97. RT-PCR analysis of cDNA from different pig cells and tissues showed that CD97 was highly expressed in leukocytes and lymph node cells. This is the first report describing the identification of a member of the EGF-TM7 family in the pig. 相似文献