共查询到20条相似文献，搜索用时 31 毫秒
1.
2.
Davis EG Sang Y Rush B Zhang G Blecha F 《Veterinary immunology and immunopathology》2005,105(1-2):163-169
NK-lysin is an antimicrobial peptide of cytotoxic and NK lymphocytes that has powerful antibacterial properties as well as antitumoral activity. Here we report the full-length cDNA and deduced amino acid sequence for equine NK-lysin. Equine NK-lysin is 67% identical to porcine NK-lysin, 53% identical to bovine NK-lysin and 41% identical to granulysin in amino acid sequence. Complete conservation of cysteine residues between equine, bovine and porcine NK-lysin suggests similar disulfide bonding patterns among these peptides. Equine NK-lysin has the most positive surface charge when compared with other homologues. Similar to expression profiles in other species, equine NK-lysin is constitutively transcribed in various lymphocytes that include CD4+ and CD8+ staining cells. These findings suggest that equine NK-lysin, similar in cDNA sequence to the porcine, bovine and human homologues may play a role in antimicrobial defense. 相似文献
3.
Swiderski CE Sobol G Lunn DP Horohov DW 《Veterinary immunology and immunopathology》2000,77(3-4):213-220
Equine interleukin-6 (IL-6) cDNA was amplified from mitogen-stimulated equine peripheral blood mononuclear cells (PBMC) using consensus sequence primers. The 727bp amplified cDNA contains the entire coding region for equine IL-6 and includes 118 bases in the 3' non-translated region. The coding sequence translates to a protein of 208 amino acids with a predicted 28 amino acid leader sequence. The mature protein of 180 amino acids has a predicted molecular mass of 20471Da without post-translational modifications. The amino acid sequence of equine IL-6 displays between 46 and 84% similarity to other mammalian IL-6 sequences. Expression of equine IL-6 in Chinese hamster ovary (CHO) cells yielded a supernatant that supported the proliferation of B9 cells in a dose-dependent manner. Treatment of B9 cells with an anti-IL-6 receptor antibody ablated the response to the recombinant equine IL-6. 相似文献
4.
We report the equine (Equs equs) and elk (Cervus elaphus) pituitary pre-prolactin (PRL) cDNA cloning, and their nucleotide and deduced amino acid sequences. Pre-PRL cDNA was obtained by RNA ligation mediated-rapid amplification of cDNA ends (RLM-RACE) and polymerase chain reaction (PCR). The elk pre-PRL cDNA exhibits two polymorphisms at positions 96 and 672, which are silent since they encode for the same amino acids, proline and isoleucine, respectively. We found no polymorphisms in the equine pre-PRL cDNA. The deduced amino acid sequence of the equine pre-PRL is 99% identical to the previously reported protein sequence. Pre-PRL mRNA is <1 kb in length and is highly expressed in the anterior pituitary gland, as demonstrated by Northern hybridization analysis. In summary, we cloned and sequenced the equine and elk pre-PRL cDNAs. The deduced amino acid sequence of elk and equine pre-PRL appears to be moderately conserved among other mammalian species. The polymorphic sites found in the elk cDNA could potentially be used in parentage testing and gene mapping. 相似文献
5.
Cloning and expression of equine insulin-like growth factor binding proteins in normal equine tendon
OBJECTIVES: To define a portion of the nucleotide sequences of each of the 6 insulin-like growth factor (IGF) binding proteins (IGFBPs) in horses and describe patterns of messenger RNA (mRNA) and protein expression for IGFBPs in normal equine tendons. ANIMALS: 7 horses. PROCEDURE: Total RNA was extracted from the tensile region of normal superficial digital flexor tendons and reverse transcribed into complimentary DNA (cDNA). The cDNA was amplified via PCR, and products representing portions of each IGFBP were cloned and sequenced. Nucleotide sequences were used to deduce the amino acid sequences, and both nucleotide and predicted amino acid sequences were compared with those published for bovine, human, mouse, and ovine IGFBPs. Gene expression was quantitated by real-time PCR assay, and protein expression was evaluated by western ligand blot (WLB). RESULTS: Clones ranged in size from 262 to 522 bp and had high degrees of sequence homology with other mammalian species. Sequence homology was highest between bovine and equine IGFBPs (86% to 95%) and amongst the IGFBP-5 sequences from the various species (92% to 95%). Message for IGFBP-2 to -6, but not IGFBP-1, was expressed in normal tendon. Protein expression for IGFBP-2, -3, and -4 was detected byWLB in normal tendon and markedly increased in damaged tendons. CONCLUSIONS AND CLINICAL RELEVANCE: Results provide basic information and tools needed for further characterization of the role of the IGF system in tendon healing and may lead to the ability to potentiate the response of healing tendon to exogenous IGF-I via concurrent manipulation of IGFBPs. 相似文献
6.
猪血清白蛋白基因的克隆与序列分析 总被引：1，自引：0，他引：1
血清白蛋白融合技术是一种开发长效重组蛋白药物的新技术.为了获得猪血清白蛋白(PSA)基因,根据GenBank(登录号:AY663543)中PSA的基因序列,设计一对引物,采用RT-PCR从新鲜猪肝组织中扩增出PSA基因的全长cDNA,产物纯化后连接至pBS-T载体,转化大肠埃希菌TG1,采用PCR法及酶切鉴定法筛选出阳性菌株后进行测序.序列分析结果表明,成功克隆了PSA基因的1 821 bp全长cDNA,与 GenBank中参照基因序列同源性为99.6%,氨基酸序列同源性为99.5%.本研究数据已提交至GenBank,并申请到一个登录号EF202601. 相似文献
7.
Donnelly BP Nixon AJ Haupt JL Dahlgren LA 《American journal of veterinary research》2006,67(7):1218-1225
OBJECTIVE: To characterize the nucleotide sequence of equine platelet-derived growth factor (PDGF)-A and -B and analyze temporal expression of these genes in equine tendon after induced tendinitis injury. Animals-18 mature horses. PROCEDURES: Genes for equine PDGF-A and -B were reverse transcribed and sequenced from synovial tissue mRNA obtained from a 3-year-old horse. Collagenase-induced lesions were created in the tensile region of the superficial digital flexor tendon in 14 horses; 3 horses served as uninjured control animals. Tendons were harvested and total RNA was isolated from experimental horses 1, 2, 4, 8, and 24 weeks after collagenase injection. Temporal gene expression for PDGF-A and -B was determined by use of quantitative PCR analysis. RESULTS: Equine PDGF-A shared 83.8% sequence and 87.5% peptide homology with human PDGF-A, with a discrepancy of 70 bp from the human sequence. Equine PDGF-B was similar in length to the human gene, sharing 90.3% and 91.7% nucleotide and peptide identity, respectively. Expression of PDGF-A mRNA in collagenase-induced tendinitis lesions was unchanged, compared with expression for normal control tendon, and remained steady throughout the 24-week study. Expression of PDGF-B mRNA decreased over time, and the expression at 24 weeks was significantly reduced, compared with expression in normal and acutely injured tendon. CONCLUSIONS AND CLINICAL RELEVANCE: Injured tendon mounts a minimal constitutive PDGF-A or -B mRNA response. Serial exogenous treatment with either PDGF isoform within the first 2 to 4 weeks after tendon injury may bolster the meager PDGF paracrine-autocrine intrinsic response to injury. 相似文献
8.
Kirisawa R Hashimoto N Tazaki M Yamanaka H Ishii R Hagiwara K Iwai H 《Veterinary immunology and immunopathology》2006,109(3-4):219-231
cDNA generated from lipopolysaccharide-stimulated equine peripheral blood mononuclear cells was used to amplify and clone type I and type II equine interleukin-1 receptors (IL-1RI and IL-1RII) using primers derived from semi-conserved regions between human and mouse IL-1RI and IL-1RII sequences, respectively. 5' and 3' terminal sequences of equine IL-1RI and IL-1RII were amplified by 5' and 3' rapid amplification of cDNA ends. The deduced amino acid sequence of equine IL-1RI demonstrated 77, 64 and 63% similarity with human, mouse and rat sequences, respectively. The predicted amino acid sequence of equine IL-1RII demonstrated 70, 60 and 58% similarity with human, mouse and rat sequences, respectively. Recombinant equine soluble IL-1RI and IL-1RII produced in insect cells bound recombinant equine IL-1alpha and IL-1beta. Furthermore, both receptors suppressed the growth inhibitory activities of equine IL-1alpha and IL-1beta toward A375 cells in a dose-dependent manner, indicating that the present equine IL-1RI and IL-1RII cDNA encodes biologically active proteins. 相似文献
9.
Watanabe M Tateyama S Togashi T Uchida K Yamaguchi R Shimizu T Sugano S 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2000,62(11):1217-1219
The nucleotide sequence of canine alpha-lactalbumin cDNA from canine mammary tissue was determined by polymerase chain reaction with degenerate primers. A 742 base pairs nucleotide sequence cloned was similar to the size of mRNA in Northern blot analysis. The cDNA encodes 142 amino acid residues containing the conserved sequence motif of alpha-lactalbumin, demonstrating the highest homology with pig (73% identity-82% similarity) among the known amino acid sequences of alpha-lactalbumin. The canine cDNA also showed 71% identity-78% similarity with human, 58-73% with mouse, 60-74% with rat, 67-77% with goat, 66-77% with cattle, and 67-76% with sheep, respectively. 相似文献
10.
OBJECTIVE: To determine the nucleotide sequence of the alphaIIb gene from canine platelet-derived cDNA. ANIMALS: 3 adult dogs. PROCEDURE: First-strand cDNA was prepared from total RNA isolated from canine platelets. The cDNA was amplified, using specific primers in polymerase chain reaction (PCR), and the nucleotide sequence was obtained from purified PCR products. RESULTS: Except for the nucleotide at position 694, results of all sequencing reactions of alphaIIb were identical for canine platelet-derived cDNA. Canine alphaIIb had 3 fewer codons than alphaIIb of humans. The nucleotide and deduced amino acid sequences of full-length canine alphaIIb shared > or = 83% similarity with the sequences established for humans. Segments of canine alphaIIb nucleotide and deduced amino acid sequences were > or = 78% similar to alphaIIb associated with 7 functional domains (extracellular, transmembrane, cytoplasmic, and 4 calcium-binding domains) in humans, with the highest degree of similarity correlating with the sequences of the 4 calcium-binding domains. Amino acid residues associated with development of alloantibodies in humans (Met837, Val837, Ile843, Ser843) are not encoded by canine alphaIIb. CONCLUSIONS AND CLINICAL RELEVANCE: The nucleotide variation at position 694 of canine alphaIIb may represent a polymorphism. The species differences in the alphaIIb sequence may contribute to variations in receptor-li gand interactions. The high degree of alphaIIb sequence conservation of the 4 calcium-binding domains implies functional importance. Some disorders associated with alphaIIbbeta3 in dogs are clinically analogous to diseases in humans, and results indicate that dogs are an appropriate model for the evaluation of gene therapy and other treatments of platelet-associated disorders. 相似文献
11.
Toribio RE Kohn CW Chew DJ Capen CC Rosol TJ 《American journal of veterinary research》2002,63(2):194-197
OBJECTIVES: To clone and sequence the cDNA for feline preproparathyroid hormone (preproPTH) and to compare that sequence with other known parathyroid hormone (PTH) sequences. SAMPLE POPULATION: Parathyroid glands from 1 healthy cat. PROCEDURES: A cDNA library was constructed in lambda phage from feline parathyroid gland mRNA and screened with a radiolabeled canine PTH probe. Positive clones were sequenced, and nucleic acid and deduced amino acid sequences were analyzed and compared with known preproPTH and PTH sequences. RESULTS: Screening of approximately 2 X 10(5) recombinant plaques revealed 3 that hybridized with the canine PTH probe; 2 clones comprised the complete sequence for feline preproPTH. Feline preproPTH cDNA consisted of a 63-base pair (bp) 5'-untranslated region (UTR), a 348-bp coding region, and a 326-bp 3'-UTR. The coding region encoded a 115-amino acid peptide. Mature feline PTH consisted of 84 amino acids. Amino acid sequence analysis revealed that feline PTH was > 83% identical to canine, bovine, swine, equine, human, and macaque PTH and 69, 71, and 44% identical to mouse, rat, and chicken PTH, respectively. Within the region responsible for hormonal activity (amino acids 1 to 34), feline PTH was > 79% identical to other mammalian PTH sequences and 64% identical to the chicken sequence. CONCLUSIONS AND CLINICAL RELEVANCE: The amino acid sequence of PTH is conserved among mammalian species. Knowledge of the cDNA sequence for feline PTH may be useful to investigate disturbances of calcium metabolism and alterations in PTH expression in cats. 相似文献
12.
Sato F Hasegawa T Katayama Y Iwanaga T Yanaihara N Kanno T Ishida N 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2000,62(9):953-959
Chromogranin A (CGA) is a member of a family of highly acidic proteins co-stored and co-released with catecholamines in the adrenal medullary cells as well as in other neurons and paraneurons. The nucleotide sequence encoding equine CGA was determined using RT-PCR and rapid amplification of complementary DNA (cDNA) ends (RACE) techniques. A total 1,828 bp of the nucleotide sequence reveals that equine CGA is a 448-residue protein preceded by an 18-residue signal peptide. Comparison of the amino acid sequence of equine CGA with those of human, porcine, bovine, mouse, rat and frog CGA showed high conservation at the NH2-terminal 1-77 amino acids regions (94.8%, 93.5%, 92.2%, 81.8%, 83.1% and 66.2%, respectively) and COOH-terminal 314-430 amino acids regions (90.6%, 81.4%, 90.6%, 80.5%, 83.3% and 39.0%, respectively), as well as a potential dibasic cleavage site, whereas the middle portion showed marked sequence variation (52.5%, 49.1%, 38.9%, 26.6%, 27.9% and 6.2%, respectively). Northern blot analysis and RT-PCR elucidated the tissue distribution of equine CGA mRNA. Its expression was confirmed not only in the adrenal medullary cells but also in other organs (cerebrum, cerebellum, pituitary gland, spinal cord, liver, thyroid gland, striated muscle, lung, spleen, kidney, parotid gland and sublingual gland). Further, in adrenal chromaffin cells and pituitary cells of the anterior-intermediate lobe, the expression was confirmed by in situ hybridization with anti-sense CGA cRNA probe. 相似文献
13.
Dannielle Tompkins Edward Hudgens David Horohov Cynthia L. Baldwin 《Veterinary immunology and immunopathology》2010,133(2-4):309-313
This report describes the initial cloning and characterization of the equine interleukin-17 (IL-17) expressed gene sequence from mRNA obtained from equine intestinal tissue and interleukin-23 (IL-23) expressed gene sequence from mRNA obtained from equine peripheral blood mononuclear cells. Equine IL-17 has 462 nucleotides in the translated region, determined by homology with known human and mouse sequences, and shares 84% and 75% identity, respectively. For the deduced amino acid sequences, the identity with human and mouse is 76% and 70%. Equine IL-23 has 579 nucleotides in the translated region. Homology with known human and mouse sequences was determined to be 89% and 77%. Deduced amino acid identities are 89% with the human sequence and 70% with the mouse sequence. The gene sequences were identified as part of the U.S. Veterinary Immune Reagent Network with a goal of developing reagents in order to aid veterinary researchers in the investigation of diseases in livestock species. 相似文献
14.
半胱亚磺酸脱羧酶(CSD)基因在大鼠睾丸组织中的表达及其序列分析 总被引：1，自引：0，他引：1
试验根据大鼠肝脏牛磺酸生物合成关键酶——半胱亚磺酸脱羧酶(CSD)基因已知核苷酸序列,设计了1对特异的寡核苷酸引物,对提取的睾丸组织总RNA进行RT-PCR扩增,扩增出468 bp的DNA片段,在国际上首次证明CSD基因在大鼠睾丸组织中存在表达,即牛磺酸可在大鼠睾丸组织中生物合成。测定了扩增的睾丸组织CSD基因核苷酸序列,并与已知肝脏CSD基因核苷酸序列及其对应的氨基酸序列作了同源性分析。结果表明:睾丸组织CSD基因核苷酸序列同已知肝脏CSD基因核苷酸序列的同源性为99.8%,其对应氨基酸序列的同源性为100%。 相似文献
15.
Molecular cloning and sequencing of equine interleukin 4 总被引：3，自引：0，他引：3
Evie V. Vandergrifft Cyprianna E. Swiderski David W. Horohov 《Veterinary immunology and immunopathology》1994,40(4):379-384
We have cloned equine interleukin 4 (IL-4) cDNA using the polymerase chain reaction (PCR) and primers based on the human IL-4 sequence. The cDNA was amplified from mitogen-stimulated equine peripheral blood mononuclear cells (PBMC). The cloned PCR product shares extensive homology with IL-4 sequences from other species. 相似文献
16.
17.
根据GenBank中公布的普通牛Hepcidin基因序列(登录号为XM-589792.3)设计1对特异性引物。采用RT-PCR技术从牦牛肝脏组织总RNA中扩增出Hepcidin基因的编码序列并进行测序分析,同时构建Hepcidin蛋白物种进化树。结果显示,经克隆获得牦牛Hepcidin基因322 bp的cDNA序列（GenBank登录号为EU863791）,含289 bp的开放阅读框,编码82个氨基酸,包括22个氨基酸的信号肽;预测Hepcidin蛋白分子质量和等电点分别为8.88 ku 和 9.24;与已知普通牛Hepcidin序列的同源性达 99%,不同物种Hepcidin蛋白具有高度的保守性,Hepcidin蛋白物种进化树符合物种进化规律。本研究为Hepcidin基因cDNA 全长克隆及基因功能研究奠定了重要基础。 相似文献
18.
为了研究白细胞介素-18(IL-18)的活性和功能，将犬脾细胞经ConA诱导，用TRIzol裂解，提取总RNA，通过反转录聚合酶链反应(RT-PCR)扩增出犬IL-18的cDNA，然后连接到pMD18-T载体上，转化DH5a感受态细胞，获得阳性重组质粒。核苷酸序列结果表明，我们得到了长686bp的基因，含有一个582bp的开放阅读框架，编码193个氨基酸。与已知犬序列同源性可高达99．6％，与已知猪序列的同源性为89．8％，所克隆基因为今后进一步研究IL-18基因的功能奠定了基础。 相似文献
19.
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