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1.
Hepcidin, a novel gene encoded, 25 residue, 2–3 KDa cysteine rich cationic peptide synthesized in liver cells play an important
role in iron metabolism in addition to its antimicrobial activity. In this study hepcidin cDNA expressed on hepatocytes of
Bubalus bubalis has been characterized and the antibacterial activity of buffalo hepcidin analog has been determined. The complete buffalo
hepcidin sequence is of 324 bp and have an ORF of 249 nucleotides from 14 to 262. The sequence analysis at nucleotide level
showed homology of 98.0% with cattle, 84.3% with pig, whereas with human, chimpanzee and gorilla 80.3%, 80.7% and 81.5% respectively.
The percent identity of buffalo hepcidin cDNA to that of japanese macaque, dog, gibbon, mouse and norway rat are81.9%, 79.9%,
79.9%, 67.9% and 65.9% respectively. The hepcidin cDNA of buffalo and cattle showed a total difference of five nucleotides
at position 15, 108, 225, 240 and 246 with no difference in deduced amino acid sequence. The base A15, C108, A225, C240, A246 in cattle are replaced by G15, G108, G225, T240 and G246 respectively in buffalo. The deduced amino acid sequence of buffalo preprohepcidin consists of 82 amino acids containing
10 strongly basic, 3 strongly acidic, 27 hydrophobic and 29 polar amino acids. The predicted molecular weight and isoelectric
point (PI) for the peptide is 8883.67 Daltons and 8.804 respectively. Phylogenetic analysis both at nucleotide and amino acid level
indicate that buffalo and cattle hepcidin sequences comprises one clad which is more closely related to pig compared to human.
The chemically synthesized analog of buffalo hepcidin using Fmoc chemistry in solid phase had antibacterial activity. 相似文献
2.
为了获得梅花鹿β-防御素-1（sika deer β-defensin-1,siBD-1）cDNA全序列,本试验以梅花鹿舌黏膜组织内提取的总RNA为模板,根据前期已获得的siBD-1 cDNA的已知部分序列设计引物,采用5'-RACE和3'-RACE技术分别扩增5'-和3'-末端序列,将此扩增产物克隆入pMD18-T载体,进行PCR、双酶切鉴定及序列测定与分析。结果表明,成功克隆出长度约为172和299 bp的siBD-1 cDNA 5'-和3'-末端序列,从而得到418 bp的siBD-1 cDNA全序列（GenBank登录号：HM588696.1）,其中包含89 bp 5'-非翻译区（UTR）、192 bp的开放阅读框（ORF）、终止密码子TAA、118 bp的3'-UTR和Poly（A）16。同源性比对结果显示,siBD-1 cDNA与水牛的肠防御素（BEBD）同源性最高,为90.6%,与牛（EBD、LAP、TAP、BNBD-4）、山羊（GBD-1、GBD-2）、驯鹿（reBD-1）、绵羊（sBD-1、sBD-2）和骆驼（caBD-1）的防御素cDNA的同源性较高,分别为83.2%、83.1%、87.3%、87.0%、87.5%、87.5%、84.4%、79.9%、77.1%和70.5%;与马（hoBD-1）和猪（pBD-1）的同源性较低,为60.3%和72.4%;而与人（hBD-2）的同源性最低,为16.0%。siBD-1成熟肽由38个氨基酸残基组成,其中包含9个带正电荷的氨基酸残基。 相似文献
3.
参照GenBank登录的绵羊凝乳酶原前体cDNA序列设计引物,以新生5d的西农萨能羊皱胃组织总RNA为模版,通过RT-PCR方法获得凝乳酶原前体cDNA,克隆测序后进行序列比对。结果表明,克隆基因为B型凝乳酶,该基因cDNA具有1292个碱基,编码381个氨基酸,包含16个氨基酸的信号肽序列和42个氨基酸的酶原序列。将其与已报道的山羊、绵羊和牛的凝乳酶原前体序列进行比对,发现核苷酸同源性分别为99.41%、98.74%和95.29%,氨基酸同源性为99.21%、98.42%和93.70%。 相似文献
4.
本研究为证明荷斯坦奶牛乳腺β-防御素的表达,扩增出6种β-防御素序列进行氨基酸分析,结果发现,扩增的6个保守的半胱氨酸,其核酸序列与NCBI数据库序列进行BLAST比对,结果LAP、BNBD4同源性100%,BNBD5为99.6%,EBD为99.5%, BNBD7、TAP为98.5%;实时荧光定量PCR检测6种防御素的mRNA水平的表达,它们之间的表达量都有显著性差异(BNBD4与BNBD5除外),其中LAP表达量最多,BNBD7,EBD,BNBD5,BNBD4次之,TAP的表达量最低。 相似文献
5.
Sequence and topological characterization of Toll-like receptor 8 gene of Indian riverine buffalo (Bubalus bubalis) 总被引：1，自引：1，他引：0
Praveen Kumar Dubey Shubham Goyal Jigyasa Aggarwal Suresh Kumar Gahlawat Periasamy Kathiravan Bishnu Prasad Mishra Ranjit Singh Kataria 《Tropical animal health and production》2012,45(1):91-99
In this study, buffalo (Bubalus bubalis) Toll-like receptor 8 (TLR8) gene has been characterized by sequence analysis and detecting polymorphism. Complete ORF of buffalo TLR8 gene was amplified using the RNA isolated from spleen tissue, which was found to be 3,102 nucleotides long encoding a 1,033 amino acid protein. Buffalo TLR8 had 10 nucleotide changes as compared to other livestock species resulting in six unique amino acid changes, four of them lying within leucine-rich repeat (LRR) domains. As compared to cattle (Bos indicus and Bos taurus), out of fifteen cysteine residues, fourteen were conserved and Cys at position 521 was replaced by Arg. Nine of the LRR domains had no amino acid change as compared to cattle, whereas LRR-C-terminus had maximum, five amino acid changes. Sequence characterization of 12 riverine and swamp buffaloes revealed presence of four polymorphic nucleotides, two of them were non-synonymous, one synonymous and one site in 3′UTR. PCR-RFLP genotyping of non-synonymous SNP 2758A>G (ILeu920Val) in Toll–interleukin-1 receptor domain of 463 swamp and riverine buffaloes showed a higher frequency of allele A in swamp (95 %) as compared to riverine (9.84 %) buffaloes. 相似文献
6.
Identification of genes of importance regarding production traits in buffalo is impaired by a paucity of genomic resources. Choice to fill this gap is to exploit data available for cow. The cross‐species application of comparative genomics tools is potential gear to investigate the buffalo genome. However, this is dependent on nucleotide sequences similarity. In this study, gene diversity between buffalo and cattle was determined using 86 gene orthologues. There was approximately 3% difference in all genes in terms of nucleotide diversity and 0.267 ± 0.134 in amino acids, indicating the possibility for successfully using cross‐species strategies for genomic studies. There were significantly higher non‐synonymous substitutions both in cattle and buffalo; however, there was similar difference in terms of dN– dS (4.414 versus 4.745) in buffalo and cattle, respectively. Higher rate of non‐synonymous substitutions at similar level in buffalo and cattle indicated a similar positive selection pressure. Results for relative rate test were assessed with the chi‐squared test. There was no significance difference on unique mutations between cattle and buffalo lineages at synonymous sites. However, there was a significance difference on unique mutations for non‐synonymous sites, indicating ongoing mutagenic process that generates substitutional mutation at approximately the same rate at silent sites. Moreover, despite of common ancestry, our results indicate a different divergent time among genes of cattle and buffalo. This is the first demonstration that variable rates of molecular evolution may be present within the family Bovidae. 相似文献
7.
本试验旨在对水牛黑色素皮质素1受体(MC1R)基因进行克隆、生物信息学分析及表达模式研究。参考牛MC1R基因(GenBank登录号:JN123363.1)序列设计引物,以本地沼泽水牛、白沼泽水牛、摩拉水牛和黄牛基因组DNA为模板,应用PCR方法扩增克隆MC1R基因片段并进行测序分析。运用QRT-PCR方法检测摩拉水牛、沼泽水牛、白沼泽水牛和黄牛皮肤组织中MC1R基因的表达模式,并通过Western blotting方法检测沼泽水牛和白沼泽水牛MC1R基因的蛋白表达差异。结果表明,应用PCR方法成功克隆了水牛MC1R基因,其编码区全长954 bp,共编码317个氨基酸。测序分析后发现沼泽水牛、白沼泽水牛、摩拉水牛和黄牛MC1R基因的核苷酸序列和氨基酸序列相似性很高。沼泽水牛与白沼泽水牛在476、618、881、930和931 bp位点上分别发生T→C、G→C、G→A、G→A和A→G突变,导致了沼泽水牛和白沼泽水牛第159位氨基酸由丝氨酸变成苯丙氨酸,第310位氨基酸由谷氨酸变成丙氨酸,第294位氨基酸由天冬氨酸变成丙氨酸,发生了非同义突变。QRT-PCR结果发现,MC1R基因在摩拉水牛、沼泽水牛和黄牛皮肤组织中的相对表达量均显著高于白沼泽水牛(P<0.05);Western blotting分析结果显示,沼泽水牛皮肤组织中MC1R蛋白的表达量高于白沼泽水牛。综上所述,白沼泽水牛MC1R基因的编码区发生氨基酸位点突变,且相对表达量和蛋白表达量均低于沼泽水牛,推测此为白沼泽水牛体内合成的黑色素缺失而导致毛色白化的主因。 相似文献
8.
LIN Yu-shu JIANG Heng-jie YANG Feng-shuo CUI Kui-qing SHI De-shun LIU Qing-you 《中国畜牧兽医》2015,42(10):2529-2537
The aim of this study was to clone and analyze the expression pattern of buffalo MC1R gene.A pair of specific primers was designed according bovine MC1R sequence (GenBank accession No.:JN123363.1),with genome DNA of swamp buffalo,White swamp buffalo,Murrah buffalo and Yellow cattle as template,MC1R was amplified by PCR.Then relative expression level of MC1R gene of swamp buffalo,White swamp buffalo,Murrah buffalo and Yellow cattle was analyzed using QRT-PCR,and protein expression was detected by Western blotting method.The results showed that the 954 bp coding region of buffalo MC1R gene was successfully cloned and sequenced,which code for 317 amino acids.The MC1R gene nucleotide sequences and amino acid sequences of swamp buffalo,White swamp buffalo,Murrah buffalo and Yellow cattle were highly conserved.Five polymorphic sites were found between White swamp buffalo and swamp buffalo of MC1R gene,including 476 T→C,618 G→C,881 G→A,930 G→A and 931 A→G,which caused three nonsynonymous mutation sites of Phe159Ser,Glu310Ala and Asp294Ala.The QRT-PCR result showed that relative expressions of MC1R gene of Murrah buffalo,swamp buffalo and Yellow cattle were significant higher than that of White swamp buffalo (P<0.05).The Western blotting results revealed that MC1R protein expression level in swamp buffalo was higher than that of White swamp buffalo.In conclusion,there were amino acids mutation in White swamp buffalo MC1R gene,and MC1R gene relative expression of White swamp buffalo was lower than that of other buffalo,which was the main reason of lacking of melanin production in White swamp buffalo. 相似文献
9.
10.
Young L 《Veterinary immunology and immunopathology》2011,140(3-4):335-340
Very few cytokines that are important to the understanding of T helper cell function are characterized in marsupials. Expression of a 645 bp cDNA product that codes for a predicted Interleukin-4 peptide of 157 amino acids was detected in the lymph node tissues of Macropus eugenii, the tammar wallaby. Using Rapid Amplification of cDNA Ends, both 5'- and 3'-untranslated regions were identified and a polyadenylation signal and three mRNA instability motifs associated with secreted cytokine molecules were also present. The translated cDNA sequence has a putative signal peptide of 24 amino acids, a predicted secondary structure that is consistent with the short-chain alpha-helical cytokine family and 82% conservation of residues associated with the Interleukin-4 family sequence motif. Comparisons of wallaby nucleotide and predicted peptide sequences with the coding domains of other vertebrate species demonstrate the diversity within this gene family; with nucleotide and amino acid identities of 74% and 59% with opossum, 52% and 32% with human and 38% and 19% with chicken homologues respectively. Despite these differences in sequence conservation, the putative Macropus eugenii Interleukin-4 mature peptide contains conserved structural motifs and predicted receptor-binding residues that suggest that it may retain functional properties associated with this important Th2 cytokine in other mammals. 相似文献
11.
Ables GP Nishibori M Kanemaki M Watanabe T 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2002,64(11):1081-1083
The natural resistance associated macrophage protein 1 (Nramp1) has been reported to confer resistance or susceptibility to Mycobacterium bovis, Salmonella typhimurium, and Leishmania donovani in the mouse, Mus musculus. A Gly and Asp substitution at position 169 of the mouse Nramp protein is invariably associated with the resistant and susceptible phenotypes, respectively. The present study aimed to detect polymorphisms in the NRAMP1 gene from different cattle and buffalo breeds. Genomic DNAs from five breeds of cattle and four breeds of buffalo were used in the study. Sequencing showed two nucleotide substitutions found in intron 4, three in exon V, and ten in intron 5. An amino acid substitution was observed at nucleotide position 1202 in exon V of the Japanese black, Angus, Philippine and Bangladesh swamp-type buffaloes which coded for Thr, while the Korean cattle, Holstein, African N'dama, Indonesian swamp-type buffalo and the Bangladesh river-type buffalo had Ile. All the breeds of cattle and buffaloes tested in this study coded for Gly at the position in exon VI which corresponds to the same amino acid of the murine Nramp1-resistant phenotype at position 169. The phylogenetic relationship among the different breeds showed a cluster comprised mainly of cattle and another one mainly of buffaloes. 相似文献
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13.
为揭示甲状旁腺激素样激素（parathyroid hormone-like hormone,PTHLH）基因对水牛繁殖性能的影响,本研究对水牛PTHLH基因进行克隆,并对其核苷酸和氨基酸序列进行生物信息学分析。以牛PTHLH基因为种子序列（GenBank登录号：NM_001290949）,应用CE Design软件设计引物序列,运用PCR扩增和测序技术获得水牛完整编码区序列,使用DNAMAN、ProtParam、SOPMA、PSORTⅡ Prediction等在线软件分析PTHLH蛋白的一级结构、二级结构、三级结构与理化性质,并进行同源性比对分析及系统进化树构建。结果显示,试验克隆了水牛PTHLH基因完整编码区序列,该序列长为534 bp,可编码177个氨基酸。水牛PTHLH基因编码区核苷酸序列与黄牛、猪、马、山羊、绵羊和骆驼的同源性分别为98.3%、90.4%、90.1%、98.1%、97.5%和89.2%,物种之间同源性较高,系统进化树分析结果与其亲缘关系远近一致,表明水牛PTHLH基因编码区在进化过程中比较保守。蛋白理化性质分析显示,水牛PTHLH蛋白分子式为C895H1451N271O266S2,分子质量为2 885 u,半衰期为30 h,理论等电点（pI）为10.00,水溶液在280 nm处的消光系数为23 950,肽链N端为蛋氨酸（Met）,不稳定系数为60.04,属于碱性不稳定蛋白;脂肪系数为72.15,总平均亲水性为－0.928,该蛋白属于不可溶性蛋白,亚细胞定位于细胞核、细胞质和线粒体。结构域预测结果显示,水牛PTHLH蛋白包含有1个PTH区域,同时还包含有1个低复杂度区域。二级结构分析显示,水牛PTHLH蛋白包含83个α-螺旋（46.89%）、17个延伸链（9.60%）、10个β-转角（5.66%）和67个无规则卷曲（37.85%）,与三级结构预测结果相一致。试验构建了PTHLH基因真核表达载体pcDNA3.1-PTHLH,并通过电泳和测序验证了载体的准确性。PTHLH基因的成功克隆及其真核表达载体的成功构建为今后研究水牛PTHLH基因的功能和遗传特性提供了材料。 相似文献
14.
N Arunmozhi SK Singh T Sarath SK Agarwal A Doiphode U Shankar 《Reproduction in domestic animals》2014,49(5):E56-E59
Buffaloes are known for their productivity as compared to average yielding cows due to higher fat percentage, better feed conversion ability and disease resistance. On the other hand, the reproductive performances of buffaloes are often considered as poor owing to late sexual maturity, weak/silent oestrus, repeat breeder and prolonged intercalving interval. The study of cascade of events during oestrus and oestrous cycle can be useful for the improvement of reproductive efficiency of buffaloes. More precisely, the hormonal changes initiated at the molecular level within the animal determine the reproductive nature of the species. Nucleotide/protein sequence analysis serves as a vital tool in analysing the binding of the hormones for their effect or functions. In this study, we have reported cloning and characterization of the complete coding (cDNA) sequence of oxytocin receptor gene (OXTR) in buffaloes. Buffalo OXTR gene contains an uninterrupted ORF of 1176 nucleotides corresponding to an inferred polypeptide length of 391 amino acids (aa). The molecular weight of the deduced aa sequence was found to be 43 kDa with an isoelectric point of 9.253 and 16.328 charge at pH 7.0. The deduced protein sequence consists of 38 strongly basic (+) (K,R), 22 strongly acidic (?) (D,E), 186 hydrophobic (A, I, L, F, W, V) and 95 Polar (N, C, Q, S, T, Y) aa. Results indicated that aspartate (D) at aa position 85 and D, R and C at aa positions 136, 137 and 138, respectively, are conserved in buffaloes. The buffalo OXTR gene shared a per cent similarity ranging from 84.7 to 98.1 and 88.5 to 97.7 at nucleotide and deduced aa sequence levels, respectively, with that of other species. Phylogram constructed on the basis of either nucleotide or deduced aa sequences of buffalo OXTR gene showed that buffalo, cattle and sheep have diverged from human and swine and formed a separate clad. The buffalo sequence has shown maximum similarity and closeness with cattle followed by sheep both at nucleotide and at aa level. 相似文献
15.
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
16.
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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18.
试验旨在克隆获得水牛脂素1（LPIN1）基因，并对其进行生物信息学分析，为揭示该基因在水牛脂肪沉积、生殖发育和泌乳调控中的作用奠定基础。本研究以水牛卵巢组织cDNA为模板，PCR扩增获得了LPIN1基因CDS区全长后测序，并结合生物信息学分析方法预测及分析蛋白质理化性质、二级结构及三级结构等。结果表明，水牛LPIN1基因编码区长2 793 bp，编码930个氨基酸。MegAlign软件分析显示，水牛LPIN1基因核苷酸序列与水牛（预测）、牦牛、黄牛、山羊、藏羚羊、绵羊、猪、骆驼、人和小鼠LPIN1基因的同源性分别为99.6%、97.9%、97.7%、97.5%、97.4%、97.1%、89.9%、89.8%、86.2%和83.5%；水牛lipin1蛋白氨基酸序列与黄牛、牦牛、山羊、藏羚羊、骆驼、猪及人的同源性分别为99%、99%、99%、99%、94%、94%及90%。应用Mega 5.0软件构建系统进化树发现，水牛与黄牛的亲缘关系最近，其次为绵羊和山羊，LPIN1基因在不同物种及进化的过程中具有高度保守性。对lipin1蛋白分析发现，其二级结构由α-螺旋、β-折叠、T-转角和无规则卷曲组成；蛋白呈弱酸性，无信号肽，亚细胞主要定位于细胞核中，存在Lipin_N、LNS2和AF1Q等结构域，其中Lipin_N、LNS2为保守结构域。 相似文献
19.
Itou T Yoshida Y Shoji Y Sugisawa H Endo T Sakai T 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2003,65(12):1351-1354
The bottlenose dolphin interleukin (IL)-8 cDNA was molecularly cloned. The dolphin IL-8 has an open reading frame of 303-bp encoding 101 amino acids. The homology of the amino acid sequence with that of other species was: sheep, 89.1%; cattle, 88.1%; pig, 85.1%; dog, 85.1%; horse, 79.2%; human, 74.5%; and macaque, 72.3%. The amino acid sequence suggested that dolphin IL-8 was a CXC chemokine. The recombinant dolphin IL-8 protein was recognized with anti-ovine IL-8 monoclonal antibody. 相似文献
20.
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