利用IBV重组N蛋白建立ELISA抗体检测试剂盒的研究   总被引：3，自引：0，他引：3  

王宏俊  陈福勇 《中国兽医杂志》2003,39(9):3-6

利用表达鸡传染性支气管炎病毒Gray株N蛋白的基因工程重组菌，表达IBV—N蛋白，并利用表达载体上带有组氨酸标记的特点，应用金属螯合填料将其纯化。用纯化的IBV—N蛋白作为包被抗原，成功地建立了一种检测血清中IBV抗体的间接ELISA方法。确定抗原最佳包被浓度为0．58μg／ml；被检血清的最佳稀释度为1：100；酶标二抗的工作浓度为1：1000。确定了血清样品阴、阳性临界0D值为0．14。与IDEXX公司IBV抗体检测试剂盒比较，结果表明自建ELISA试剂盒具有较高的敏感性和特异性。  相似文献   

Expression of a recombinant crustacean hyperglycemic hormone of the kuruma prawn Penaeus japonicus in methylotrophic yeast Pichia pastoris     

TSUYOSHI  OHIRA  HIDEKAZU  KATAYAMA  KATSUMI  AIDA  HIROMICHI  NAGASAWA 《Fisheries Science》2003,69(1):95-100

ABSTRACT:   Until now, six crustacean hyperglycemic hormones (CHH) designated Pej-SGP-I, -II, -III, -V, -VI and -VII have been characterized in the kuruma prawn Penaeus japonicus . All CHH consist of 72 amino acid residues and have an amidated carboxyl (C)-terminus. In the present study, we expressed Pej-SGP-III in methylotrophic yeast Pichia pastoris in order to obtain a large quantity of recombinant CHH possessing biological activity. A cDNA encoding Pej-SGP-III that had been previously cloned was processed by polymerase chain reaction (PCR) and the resulting product was ligated into an expression vector. Pichia pastoris was transformed with this vector after which a recombinant Pej-SGP-III was expressed having an additional amino acid residue (glycine) at the C-terminus (rPej-SGP-III-Gly), a form considered to be a putative precursor of this hormone. rPej-SGP-III-Gly secreted into the culture medium was purified by reversed-phase HPLC, and amidated using a peptidylglycine alpha-amidating enzyme. The amidated rPej-SGP-III (rPej-SGP-III) showed hyperglycemic activity in in vivo bioassay almost comparable to that of the natural Pej-SGP-III. rPej-SGP-III thus obtained will be a useful tool not only for its physiological study but also for the determination of its 3-D structure.  相似文献   

A review of antimicrobial peptides: defensins and related cationic peptides     

Evans EW  Harmon BG 《Veterinary clinical pathology / American Society for Veterinary Clinical Pathology》1995,24(4):109-116

Cationic antimicrobial peptides are present throughout the plant and animal kingdoms and bear striking structural and functional similarities across species lines. They provide primitive, nonspecific means of combating a variety of bacteria, fungi, enveloped viruses, and protozoa. Some are also cytotoxic against host cells, including neoplastic cells. Cationic antimicrobial peptides may play various roles in inflammation and tissue repair. Antimicrobial peptides are found in epithelial tissues regularly exposed to microbial attack as well as in cells whose primary function is defense against potential pathogens. They constitute an important part of the nonoxidative antimicrobial arsenal of leukocytes. They are preformed and/or readily synthesized when the cells are stimulated by exposure to pathogens. They exert their effects directly by inserting into membranes of target cells and forming ion channels which increase membrane permeability; however, antimicrobial peptides can also act as opsonins to facilitate phagocytosis. Resistance to defensins is a virulence factor for organisms such as Salmonella sp. The study of cationic antimicrobial peptides is increasing our understanding of innate immunity, inflammation, and the pathogenesis of genetic diseases such as specific granule disease in humans. Therapeutic applications of antimicrobial peptides are currently under investigation.  相似文献   

Evaluation of a radioimmunoassay for type III procollagen peptide in the dog     

Dimski DS  Brooks CL  Johnson SE 《Veterinary clinical pathology / American Society for Veterinary Clinical Pathology》1990,19(2):40-44

Type III procollagen peptide (P-3-P) is a serum marker for hepatic fibrosis in humans. The utility of a commercially available radioimmunoassay for P-3-P was evaluated in the dog. The specificity of the assay was assessed by polyacrylamide gel electrophoresis (PAGE) of canine serum and purified bovine P-3-P, followed by Western immunoblotting with rabbit aniti-P-3-P serum. The sensitivity was assessed by performing the radioimmunoassay on dilutions of sera from 22 dogs. Polyacrylamide gel electrophoresis of purified bovine P-3-P and sera from two dogs suspected of having elevated P-3-P concentrations revealed no homologous bands of staining. Western immunoblotting showed marked cross-reactivity of the high antisera concentrations with several components of the serum proteins, but none corresponding to the purified P-3-P. All tested sera from dogs had minimal competitive binding with radiolabeled P-3-P in the radioimmunoassay. Dilution curves of dog sera did not parallel either the standard curve or the dilution curve of a known test human serum. There were no statistically different P-3-P concentrations in any of the groups of dogs studied. It was concluded that currently available radioimmunoassay kits for the measurement of P-3-P in the human are not applicable in the dog. Seemingly, the structure or metabolism of canine P-3-P may vary significantly from that of the bovine or human, limiting the sensitivity and specificity of this assay in the dog.  相似文献   

饲粮中添加抗菌肽对肉鸭增重及血清尿素氮、总蛋白水平的影响   总被引：12，自引：1，他引：12  

陈晓生  张辉华  罗竞彪  温刘发  黄国庆  黄自然 《中国饲料》2005,(10):21-23

240只1日龄肉鸭随机成分4组,每组设3个重复,对照组用基础日粮,试验组在基础日粮上每千克分别添加1、2、3mL抗菌肽制剂。结果显示:1)2、3mL/kg添加量组增重效果显著(P<0.05),3mL/kg添加量组最佳;净肉率升高,达到显著水平(P<0.05),其中主要是胸肌率上升;腹脂率降低但差异不显著(P>0.05);料重比无显著变化(P>0.05)。2)血清总蛋白浓度差异不显著(P>0.05),但尿素氮浓度下降,2、3mL/kg添加量组达显著水平(P<0.05),3mL/kg添加量组最低。  相似文献   

反刍动物肽营养研究进展     

张香斋  李成会 《中国饲料》2005,(12):22-24

本文综述了反刍动物肽的吸收特点及影响因素,肽的吸收代谢机制,肽对瘤胃微生物的调控及对反刍动物生产性能发挥的作用,以及饲料蛋白源活性肽的开发应用。  相似文献   

Effect of CGRP on level of lung endogenous NO in rabbits with acute lung injury     

LIAO Jian-jun  LI Huan-zhang  Qi Hao-wen 《园艺学报》2003,19(2):263-265

AIM:To examine whether calcitonin gene-related peptide (CGRP) enhances nitric oxide (NO) level in pulmonary circulation blood and observe the influence of CGRP on mean pulmonary artery pressure (mPAP) in rabbits with acute lung injury (ALI) caused by oleic acid.METHODS: The level of NO was assessed by measuring the presence of nitrite in cervical artery blood by the Griess reaction, mPAP was measured with right ventricular catheter.RESULTS:The level of nitrite in cervical artery blood was significantly increased and the mPAP was markedly reduced after administration of CGRP intravenousely.CONCLUSION:CGRP enhanced the NO level of pulmonary circulation blood and reduces the mPAP significantly in rabbits with ALI.  相似文献   

Biosynthesis of Depsipeptide Mycotoxins in Fusarium     

Till Hornbogen  Mirko Glinski  Rainer Zocher 《European journal of plant pathology / European Foundation for Plant Pathology》2002,108(7):713-718

The cyclic hexadepsipeptide enniatin is known as a phytopathogenic compound from Fusaria causing necrosis and wilt. The molecule consists of three alternating residues each of a branched chain amino acid and D-hydroxyisovaleric acid (D-Hiv). Enniatins are synthesized by a 347kDa multienzyme (enniatin synthetase) via a thiol template mechanism. The corresponding gene esyn1 has an open reading frame of 9393 nucleotides and harbours two modules, one responsible for D-hydroxy acid activation and one for L-amino acid activation with an integrated N-methyltransferase domain. Such methyltransferases build an homologous group among N-methyl peptide synthetases. Enniatins are synthesized by step-wise condensation of dipeptidol building blocks in an iterative manner resembling fatty acid synthesis. A key enzyme in enniatin biosynthesis is the NADPH-dependent D-2-hydroxyisovalerate dehydrogenase, that supplies enniatin synthetase with D-Hiv. Enniatins contribute to the wilt toxic character of Fusaria. Virulence was significantly reduced in F. avenaceum after disruption of the esyn1 gene.  相似文献   

Role of polymerase β in DNA metabolism and genomic instability     

WANG Gu-liang  YU Ying-nian 《园艺学报》2002,18(4):427-432

The major role of DNA polymerase β was thought to be limited in its involvement in short patch base excision repair by removing 5’-deoxyribose phosphate and base insertion. However, the recent researches indicate that polymerase β might take part in a wide spectrum of DNA metabolism reactions, including long patch base excision repair, DNA replication, recombination, meiosis and transleisional DNA synthesis. Because of its wide and important cellular function, an inappropriate intracellular polymerase β level might be associated with genomic instability. Down-regulation or mutation of polymerase β is mutagenic due to deficient in DNA repair, while overexpression of this error-prone β polymerase might perturb the normal function of other accurate polymerases and cause genomic instability as well.  相似文献   

Purification and characterization of an antibacterial compound produced by Agrobacterium vitis strain E26 with activity against A. tumefaciens     

H. M. Wang  H. X. Wang  T. B. Ng †  J. Y. Li 《Plant pathology》2003,52(2):134-139

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