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1.
The purpose of this work was to produce rabbit anti-cockatiel immunoglobulin G (IgG) and compare its cross-reactivity with sera from eight other psittacine birds: Quaker parakeet, budgerigar, green-wing macaw, blue-fronted Amazon parrot, eclectus parrot, African grey parrot, Patagonian conure, Moluccan cockatoo. Cockatiel IgG did not bind to protein A or G; therefore, these proteins could not be used in column chromatography to isolate the IgG. A combination of serum IgG precipitation by ammonium sulfate and yolk IgG extraction from egg was loaded in sodium dodecyl sulfate-polyacrylamide gel upon which the IgG was resolved by electrophoresis. The resolved IgG in sodium dodecyl sulfate-polyacrylamide gel was stained with Coomassie blue, cut, crushed in phosphate-buffered saline, and injected into rabbits. The rabbit anti-cockatiel IgG produced in this way reacted with a single protein in gel immunodiffusion assay with all nine psittacine bird sera but not with those of chicken and ostrich. Immunoelectrophoresis confirmed the cross-reactivity of different psittacine sera with the anti-cockatiel IgG serum but not with ostrich and chicken sera. This antiserum detected antibody responses in sera from cockatiels vaccinated against chlamydial major outer membrane protein in an immunoblot assay.  相似文献   

2.
鸡Rab7b在细胞晚期内吞体定位和结合恒定链的分子特征   总被引:1,自引:1,他引:0  
Rab蛋白是介导胞内活性蛋白转运的分子家族。恒定链(invariant chain,Ii)具有协助抗原肽转运、B细胞成熟和作为细胞因子的受体等免疫学功能。前期研究发现,鸡(Gallus gallus)Rab5a在细胞早期内吞体中与Ii结合,但不清楚是否还有其他转运蛋白有该特性。基于Rab7b分子定位于晚期内吞体,并与胞内分子转运相关,本研究探索了鸡Rab7b与Ii结合的分子结构特征。首先,用自行设计的引物扩增了鸡和小鼠(Mus musculusRab7b基因,并进行了氨基酸同源性比对分析;构建了含鸡Rab7b及其67位氨基酸突变体Rab7bQ67L的原核和真核重组质粒。其次,将含红色荧光蛋白和目的基因的重组质粒转染鼠巨噬细胞系Raw264.7,培养后用绿色荧光素(FITC)标记的鼠抗晚期内吞体蛋白1(LAMP1)抗体染色,观察目的蛋白在内吞体的定位。进一步将鸡Rab7b和Rab7bQ67L分别与Ii共转染293T细胞,观察它们与Ii的共定位。最后用拉下法和免疫印迹检测鸡Rab7b及Rab7bQ67L与Ii的结合。结果表明,所克隆获得的鸡Rab7b基因与预期大小一致,其开放阅读框为624 bp,编码208个氨基酸。鸡和鼠的Rab7b蛋白质结构相似性为74%。尽管鸡Rab7b和突变体Rab7bQ67L均能结合Ii,但只有Rab7b可以定位于晚期内吞体,而突变体却改变了该定位特性。综上所述,鸡Rab7b与Ii不仅在晚期内吞体共定位,而且还能互相结合;鸡Rab7b的第67位氨基酸影响其在细胞内定位而不影响与Ii结合。综上表明,Rab分子参与了Ii在细胞内细胞器的转运,为进一步研究Ii在细胞内的转运机制提供了新的途径。  相似文献   

3.
Rab is a family of molecules that mediate the transport of active proteins in cells. The invariant chain (Ii) has the immunological functions of assisting antigen peptide transport, B cell maturation, and acting as the receptor of cytokines. Previous studies showed that Rab5a of chicken (Gallus gallus) is bound to Ii in the early endocytosis, but it is not clear whether there are other Rab molecules with this property. Based on the localization of Rab7b in late endocytosis and its relationship with intracellular molecular transport, this study explored the molecular structure of Rab7b binding to Ii. First, Rab7b genes of chicken and mouse (Mus musculus) were amplified with self-designed primers, and the amino acid homology was analyzed; the prokaryotic and eukaryotic recombinant plasmids containing Rab7b and its 67 amino acid mutant Rab7bQ67L were constructed. Secondly, the recombinant plasmid containing red fluorescent protein and target gene was transfected into mouse macrophage line Raw264.7. After culture, the recombinant plasmid was stained with FITC labeled mouse anti late endosomal protein 1 (LAMP1) antibody to observe the localization of the target protein. Furthermore, chicken Rab7b and Rab7bQ67L were co-transfected with Ii to observe their co-localization with Ii. Finally, the binding of Rab7b and its mutants to Ii were detected by pull-down technique and Western blotting. The results showed that the Rab7b gene was the same size as expected, and its open reading frame was 624 bp, encoding 208 amino acids. The homology of Rab7b protein structure between chicken and mouse was 74%. Although both Rab7b and Rab7bQ67L could bind to Ii, it was Rab7b rather than Rab7bQ67L could locate in the late endocytosis. In conclusion, Rab7b and Ii were not only co-located in the late endocytosis, but also combined with each other; the 67th amino acid of Rab7b affected its location in cells but not with Ii. These results suggest that Rab molecules are involved in the transport of Ii in intracellular organelles, which provides a new way to further study the mechanism of Ii transport in cells.  相似文献   

4.
125-I-IgG binding activities were observed with 15 (17%) of 90 S. intermedius isolates from dogs and 39 (95%) of 41 S. hyicus isolates from pigs. Binding activities were not detected with S. hyicus isolates from cows. The IgG binding proteins of 2 S. intermedius, 2 S. hyicus, and protein A from S. aureus Cowan I were isolated from their cell surfaces. The proteins precipitated with IgG preparations from human, rabbit, pig, dog and horse, but not with IgG from cow, mouse and chicken. This indicated that these IgG binding proteins could be classified as type I receptors. In addition, the isolated proteins from all 3 staphylococcal species precipitated with polyclonal chicken anti-protein A antiserum. SDS-PAGE, Western blotting and gel isoelectric focussing of the proteins revealed numerous bands in the 42,000 D range and acid isoelectric points. The isoelectric point of the isolated proteins from both S. intermedius cultures was slightly more acidic than those from S. hyicus and S. aureus. The present results indicate a close functional and antigenic similarity, if not identity, between IgG binding proteins of S. intermedius and S. hyicus, and protein A of S. aureus.  相似文献   

5.
葡萄球菌A蛋白(SPA)和链球菌G蛋白(SPG)的IgG结合区编码基因融合后,克隆到大肠杆菌表达载体PGEX,SPA/SPG融合蛋白(SPAG)得到高效表达。可溶性SPAG~GST产物可被谷胱甘肽亲和柱一次性纯化。用琼扩和ELISA试验测定了表达产物与不同种属动物血清反应的性能。SPAG—GST与所有SPA、SPG各自反应的动物IgG结合,对小鼠IgG的反应优于SPA或SPG。在所测试的23种动物血清中,融合蛋白与大部分哺乳动物Ig反应。  相似文献   

6.
Receptors for IgG1 and IgG2 (FcR) on peripheral blood lymphocytes from cows with chronic lymphocytic leukemia (CLL) were detected by their ability to bind homologous IgG1 and IgG2 in fluorometric binding assay. Scatchard plots at 4 degrees C demonstrated that IgG1 bound the same number of FcR per cell (3.12 +/- 0.69 X 10(5)) as IgG2 (2.89 +/- 0.69 X 10(5)). The receptors bound IgG2 with an affinity of 4.09 +/- 1.08 X 10(5) 1/M and IgG1 with an affinity of 2.73 +/- 0.55 X 10(5) 1/M, although the difference was not of statistical significance (0.1 greater than P greater than 0.05). Inhibition studies demonstrated that the two ligands could inhibit each other. It might be assumed that FcR for the two subclasses were identical.  相似文献   

7.
通过比较孔雀 Ig G与鸡 Ig G的相似性 ,证实鸡疾病诊断试剂用于孔雀疾病诊断的可行性。用饱和硫酸铵盐析法提纯鸡 Ig G、孔雀 Ig G,比较 Ig G粗提物的 SDS-聚丙烯酰胺凝胶电泳图谱。用直接 EL ISA方法分析孔雀 Ig G、鸡 Ig G与抗鸡 Ig G结合程度的差异 ,并用蛋白免疫转印法进一步验证二者与抗鸡 Ig G的结合情况。最后 ,应用数种鸡疾病诊断试剂盒及血凝抑制实验进行孔雀疾病的血清学调查及诊断应用初步研究。结果表明 :孔雀 Ig G与鸡 Ig G具有较大相似性 ,并能够与兔抗鸡酶标二抗结合 ,但结合力弱于鸡 Ig G;可应用鸡疾病诊断试剂盒诊断孔雀疾病  相似文献   

8.
One hundred and twenty bacterial strains were tested for non-immune binding of radiolabelled bovine, ovine, caprine and equine immunoglobulins. Bacteria possessing previously defined IgG receptors interacted in a well defined manner with purified IgG subclass immunoglobulins. Human group C and G streptococci carrying IgG receptors type III were capable of binding all IgG subclasses in the four mammalian species studied. Protein A-containing staphylococci demonstrated a restricted specificity with binding of bovine IgG1, ovine IgG1, caprine IgG1 and IgG2 as well as equine IgG(ab). Group A streptococci which can bind human IgG did not show specific reactivity. A new type of binding unrelated to the regular Fc-mediated binding was observed with equine IgG(T).The differences in specificity for IgG subclasses suggest that structures with binding capacity to streptococcal type III Fc receptors are different from staphylococcal protein A reactive sites. Inhibition experiments performed with purified immunoglobulins showed that individual IgG subclasses differed greatly in their inhibiting capacity reflecting differences in avidity.The high avidity and the broad, unrestricted immunoglobulin G reactivity of streptococcal IgG receptor type III indicate that human group C and G streptococci may provide a valuable tool for solid phase absorption of immunoglobulins from several mammalian species.  相似文献   

9.
Porcine AIDA-I positive Escherichia coli causes diarrhea in neonatal piglets and AIDA-I adhesin is an important virulence factor involved in intestinal colonization with biofilm formation. This biofilm consists of AIDA-I(+)E. coli bacteria stratified within mucus layers covering the intestinal mucosa. Based on the intimate interaction between AIDA-I(+)E. coli and mucus within the intestinal biofilm, we hypothesized that porcine intestinal mucus contains receptor(s) for AIDA-I adhesin. Since porcine AIDA-I receptors have not been identified, we employed affinity chromatography and in vitro adhesion assays to investigate AIDA-I binding proteins in porcine intestinal mucus that might serve as receptors for attachment of AIDA-I positive E. coli. We demonstrated that porcine mucus contains 65 and 120kDa proteins (p65 and p120) that bind with high affinity to purified AIDA-I adhesin and that AIDA-I positive E. coli binds to these proteins with higher affinity than do AIDA-I negative mutant. The identity of p65 was not determined based on LC-MS/MS data, whereas p120 was matched to two nuclear proteins (namely, DNA damage binding protein and splicing factor 3b) and one cytoplasmic protein, which is an IgG Fc binding protein. Based on similar amino acid homology, molecular weight, structural similarity to mucin and reported evidence of being secreted by goblet cells into the intestinal lumen, we think that the IgG Fc binding protein is most likely candidate to serve as a potential receptor in intestinal mucus for AIDA-I adhesin.  相似文献   

10.
The binding ability of staphylococcal protein A (SpA) and streptococcal protein G (SpG) to egg yolk antibodies of four fowl species (turkey, duck, moskovy duck and goose) was studied and compared with the binding ability to three serum antibodies from chicken, horse and cattle. SpA and SpG were not able to bind to any of the avian immunoglobulins.  相似文献   

11.
This study was carried out to develop and characterize mouse monoclonal antibodies (mAbs) against chicken CD25 (chCD25), the alpha chain of the interleukin-2 (IL-2) receptor. A recombinant chimeric chCD25/IgG4 fusion protein was expressed in Chinese hamster ovary (CHO) cells and isolated from spent cell culture medium by protein G affinity chromatography. Purified chCD25 protein was used to immunize mice, from which 54 stable hybridomas secreting chCD25 mAbs were produced. Two mAbs, chCD25-32 and chCD25-54, with high binding affinity for chCD25-expressing CHO cells were selected for further characterization. By flow cytometry, both mAbs detected cells in the spleen, bursa of Fabricius, intestinal duodenum, and immunostained established chicken T cell, B cell, and macrophage cell lines. Both mAbs reacted with a 55 kDa protein on Western blots of lysates from concanavalin A (Con A)-stimulated spleen mononuclear cells. Intraperitoneal injection of chickens with bacterial lipopolysaccharide increased the percentage of chCD25(+) spleen cells by approximately 4-fold compared with untreated animals. In vitro stimulation of spleen cells with Con A increased the percentage of chCD25(+) cells by up to 50-fold compared with cells treated with medium alone. Finally, the chCD25-32 mAb suppressed IL-2-driven spleen cell proliferation and reduced IL-2-induced nitric oxide production. These mAbs may be useful for future investigation of chicken regulatory T cells.  相似文献   

12.
A "double-sandwich" ELISA for the detection and measurement of a heat-labile enterotoxin produced by porcine enterotoxigenic strains of Escherichia coli (LTp) is described. In contrast with other heat-labile toxins, LTp did not bind to agarose gels and exhibited a very low affinity for GM1 in the classical GM1-ELISA technique. The similarity of LTp with cholera toxin was confirmed by immunoblotting. This property allowed the binding of LTp to rabbit IgG anti-cholera toxin antibodies (covalently linked to polystyrene plates) and sheep anti-cholera toxin serum. The immunocomplex was revealed by anti-sheep immunoglobulin antibodies conjugated with peroxidase. Application of the "double-sandwich" ELISA to the quantitation of toxin production by two strains, which differ only in the presence or the absence of the K88ab antigen, showed that the Ent+, K88+ strain produced significantly less toxin than the Ent+, K88- derivative.  相似文献   

13.
Many proteins of the calcium-dependent (C-type) lectin family have been shown to play an important role in innate immunity. They can bind to a broad range of carbohydrates, which enables them to interact with ligands present on the surface of micro-organisms. We previously reported the finding of a new putative chicken lectin, which was predominantly localized to the respiratory tract, and thus termed chicken lung lectin (cLL). In order to investigate the biochemical and biophysical properties of cLL, the recombinant protein was expressed, affinity purified and characterized. Recombinant cLL was expressed as four differently sized peptides, which is most likely due to post-translational modification. Crosslinking of the protein led to the formation of two high-molecular weight products, indicating that cLL forms trimeric and possibly even multimeric subunits. cLL was shown to have lectin activity, preferentially binding to alpha-mannose in a calcium-dependent manner. Furthermore, cLL was shown to inhibit the haemagglutination-activity of human isolates of influenza A virus, subtype H3N2 and H1N1. These result show that cLL is a true C-type lectin with a very distinct sugar specificity, and that this chicken lectin could play an important role in innate immunity.  相似文献   

14.
This study was carried out to develop and characterize mouse monoclonal antibodies (mAbs) against chicken CD83 (chCD83), a membrane-bound glycoprotein belonging to the immunoglobulin superfamily that is primarily expressed on mature dendritic cells (DCs). A recombinant chCD83/IgG4 fusion protein containing the extracellular region of chCD83 was expressed in Chinese Hamster Ovary (CHO) cells and isolated from the spent cell culture medium by protein G affinity chromatography. The extracellular region of the chCD83 protein was purified and used to immunize mice. A cell fusion was performed, from which 342 hybridomas were screened for mAbs to chCD83. Two mAbs, chCD83-159 and chCD83-227, stained the greatest percentage of chCD83-transfected CHO cells and were selected for further characterization. By flow cytometry, both mAbs reacted with a chicken macrophage cell line, HD11. Both mAbs also recognized a single 53 kDa protein on Western blots of lysates from lipopolysaccharide-stimulated spleen mononuclear cells or unstimulated HD11 cells. Immunostaining of chicken secondary lymphoid organs identified chCD83(+) cells with morphologic and subtissue localization properties comparable to mammalian DCs. In vitro stimulation of spleen mononuclear cells with concanavalin A (Con A) decreased the percentage of chCD83(+) cells compared with cells treated with medium alone. Interestingly, spleen cells treated with Con A in the presence of chCD83-227 mAb exhibited decreased percentage of MHCII(+) cells compared with cells treated with an isotype-matched negative control mAb. These chCD83 mAbs may be useful for future investigations of chicken immune cell maturation and mechanisms of action.  相似文献   

15.
Pig paramyxovirus of the blue eye disease (PPBED) is a novel member of the paramyxoviridac family which infects pigs. In neonatal pigs it causes neurological damage, whereas in adult pigs it affects the reproductive function. As PPBED damages the new-born pig central nervous system (CNS), it is important to study whether PPBED binds to the membrane proteins of all brain tissue, or selectively binds to neuronal tissue of the brain stem, olfactory bulb, hippocampus, cerebellum, frontal, temporal and parietal brain cortex. It is also important to establish whether it also infects neurones obtained from new-born, 60-day-old and adult pigs, and the role of carbohydrate residues in virus binding. The effect on virus binding of polyclonal antibodies against viral envelope proteins was also studied. Binding studies were performed using dot blot and virus overlay protein binding assays. PPBED was able to bind to membrane proteins from all brain regions, particularly to a protein band of approximately 116 kDa. Neuraminidase treatment of neuronal membrane proteins decreased virus binding; subsequent treatments with beta-galactosidase and manosidase did not increase virus binding inhibition. N-glycosidase F and trypsin also decreased virus binding, but not the O-glycanase. Antibodies against viral haemagglutinin-neuraminidase blocked virus binding more efficiently than antibodies against viral fusion protein. In conclusion: (1) PPBFD is able to bind to pig neurones of all brain regions studied and at all ages analysed; (2) a 116 kDa membrane protein containing sialic acid residues with an N-linked oligosaccharide chain was specifically recognized; (3) PPBED haemagglutinin-neuraminidase protein seems to play a central role in neural receptor recognition.  相似文献   

16.
A murine monoclonal antibody specific for bovine immunoglobulin (Ig) G2 was used to develop an enzyme-linked immunosorbent assay (ELISA) to measure serum antibodies against the bovine pathogen Staphylococcus aureus. Anion exchange chromatography was used to prepare IgG2 from serum taken from a mastitic cow infected with S aureus. Specific-IgG2 antibodies binding S aureus were purified with an affinity column, using a heat-killed, low protein A strain of S aureus (M-10). Purified antibodies did not contain IgG1 or IgM and were composed of greater than 95% heavy and light chains determined on the basis of polyacrylamide gel electrophoresis. Purified IgG2 anti-S aureus antibody was used as a reference in an ELISA that (i) could detect 5.0 ng of IgG2, (ii) was specific for IgG2 antibodies binding S aureus, (iii) was precise within and among different assays, (iv) yielded 112% recovery of the purified standard, and (v) when diluted in nonspecific IgG2, generated a curve that was parallel to the standard when a sample was serially diluted. A field study with cows having elevated California mastitis test scores showed evidence of infection with S aureus, as judged by a 59% increase (P less than 0.01) in IgG2 S aureus-specific antibodies and a 25% increase (P less than 0.05) in total IgG2 antibodies. There were no differences in IgG1 concentrations in plasma. Using indirect immunofluorescence, we also confirmed that bovine polymorphonuclear cells bound IgG2 preferentially over IgG1 or IgM. Measurement of antigen-specific IgG2 antibodies may therefore be useful as an index of specific antibody immunity to mastitis-causing organisms.  相似文献   

17.
Although the importance of carbohydrate recognition by sperm during egg zona pellucida binding has been widely reported, the sperm molecular species that recognize the carbohydrates are poorly characterized. Our previous cytochemical study indicated that two kinds of carbohydrate-binding proteins are expressed on porcine sperm heads-one recognizes N-acetyllactosamine (Galβ1-4GlcNAc-), and the other recognizes the Lewis X structure (Galβ1-4(Fucα1-3)GlcNAc-). For this report, we used proteomic techniques to characterize the sperm proteins that bind N-acetyllactosamine. Porcine sperm plasma membrane was solubilized with a detergent solution and subjected to sequential chromatography with dextran sulfate agarose, affinity, and hydroxyapatite, and the binding activities in the eluates were monitored by a solid-phase binding assay. The tryptic peptides of two proteins most likely associated with the binding activities were subjected to tandem mass spectrometry sequencing. A subsequent database search identified one of the two proteins as predicted disintegrin and metalloprotease domain-containing protein 20-like (XP_003128672). The other protein was identified as disintegrin and metalloprotease domain-containing protein 5 (AB613817) by database searches for homologous amino acid sequences, cDNA cloning, nucleotide sequencing and nucleotide database searches. Furthermore, two-dimensional blue native/SDS-PAGE demonstrated that they formed a variety of non-covalent complexes. Therefore, these ADAM complexes probably are responsible for the N-acetyllactosamine-binding activity. An affinity-purified fraction containing these ADAM complexes showed zona pellucida-binding activity, though the activity was relatively weak, and the presence of another zona pellucida-binding protein that probably works in concert with these ADAM complexes was suggested. Immunofluorescence testing suggested that ADAM20-like was localized on the anterior part of the sperm plasma membrane.  相似文献   

18.
Hybridomas producing monoclonal antibodies to Ehrlichia risticii were developed to provide a means of molecular investigation of the biochemical and immunopathologic characteristics of the organism. All of 6 stable monoclonal antibodies obtained were IgG isotypes. The ascitic fluid titers induced by the hybridomas ranged from 10(2) to 10(7). Competitive binding experiments conducted by ELISA and binding of labeled protein A to antigen-antibody complexes indicated competition among monoclonal antibodies. Two monoclonal antibodies (HybI and 14D4) were reactive in an indirect fluorescent antibody test; these antibodies also bound a maximum of labeled protein A, indicating recognition of epitopes on the surface of the ehrlichia. Protein specificity of monoclonal antibodies could not be demonstrated with western blot procedure. HybI monoclonal antibody, however, did precipitate the 28 kD protein from 125I-surface-labeled ehrlichiae and was shown to be specific to E risticii on the basis of nonreactivity with E sennetsu, using the indirect fluorescent antibody test. By use of the different monoclonal antibodies as probes, more definitive molecular studies now will be feasible.  相似文献   

19.
Quick and cost-effective serologic assays, such as those based on enzyme-linked immunosorbent assay (ELISA) technology, are useful for screening animal populations for infectious diseases. Recombinant protein G is described as an almost universal ELISA conjugate for the detection of antibodies from a wide range of animal species. However, there is limited data documenting the ability of protein G to bind immunoglobulin (Ig) from many captive and free-ranging nondomestic hoofstock (Order Artiodactyla, e.g., elk, antelope, bison). Protein G binding to Ig from 11 species within this taxonomic order (addax, antelope, bison, bontebok, elk, impala, kudu/nyala, muntjac, oryx, sheep, and white-tailed deer) and 2 control species (bovine and chicken) was assessed. A serum Ig enrichment protocol, using high-performance liquid chromatography (HPLC), was optimized in bovids (Bos taurus) and then applied to the other study species. Binding assays were performed by adding protein G to microtiter wells coated with titrated dilutions of enriched artiodactyl Ig. Optical densities were measured and binding curves generated. Differences in protein G binding were observed, both within and among species, as well as within taxonomic families. Significant intraspecies binding variation was observed for 7 species tested (antelope, oryx, sheep, muntjac, impala, bontebok, and addax). No statistically significant intraspecies differences in protein G binding were found for Ig from bison, elk, kudu/nyala, white-tailed deer, plus control species (cattle and chicken). Binding of protein G to Ig from impala, muntjac, and elk was statistically different from the positive control (cattle), with muntjac binding curves statistically comparable with the negative control (chicken). For the other 7 species tested, binding curves illustrated the ability of protein G to bind Ig as well as, or better than, the positive control. These findings expand the list of animal species whose Ig is capable of being detected using recombinant protein G, with the caveat that protein G does not bind Ig uniformly in closely related species. It is concluded that recombinant protein G conjugates may serve as useful reagents for serodiagnosis by ELISA in nondomestic hoofstock, although different assay interpretation algorithms and assay protocols may need to be developed on a per species basis for maximum diagnostic effectiveness.  相似文献   

20.
For many pathogens, adherence and/or invasion involve association with host extracellular matrix molecules, such as fibronectin (Fn). Pasteurella multocida was found to bind significantly to Fn and collagen type IX but not to laminin and collagen types IV and X. The binding of P. multocida to Fn was dose-dependent and was inhibited by heparin (Hep). Removal of polysaccharide capsule enhanced the binding capacity of the bacterium to Fn and inhibition by Hep. Protease treatment of bacteria decreased binding, implicating surface protein(s) as adhesive components. Investigation of the binding domain(s) of P. multocida on the Fn molecule revealed preferential binding to the N-terminal Hep-binding domain of Fn but not to the carboxyl-terminal Hep-binding domain. Furthermore, Fn, and anti-Fn antibodies inhibited P. multocida adherence to Madin-Darby bovine kidney cells, suggesting the involvement of Fn in the bacterium adherence to host cells. Ligand blotting, batch affinity purification and MALDI-TOF mass spectrometry implicated several proteins as putative adhesins of P. multocida in the Fn-mediated adherence. Taken together, the data suggest that P. multocida-Fn interaction may play a role in the bacterium adherence to host cells, and this may be mediated by bacterial surface proteins with preferential affinity for the Hep-1 binding domain of Fn.  相似文献   

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