We constructed fusion proteins consisting of fluorescence-enhanced green fluorescent protein (EGFP) and soluble domain of duck B-cell-activating factor of the TNF family (dsBAFF). The soluble EGFP/dsBAFF was efficiently expressed in Escherichia coli BL 21 (DE3) and was purified in milligram amounts using metal chellate affinity chromatography. The fusion protein exhibited similar fluorescence spectra with free EGFP and promoted the survival of duck bursal B cells in vitro as well as dsBAFF. EGFP/dsBAFF has shown specific binding to duck BAFF receptors positive-cells and the stained cells could be analyzed with flow cytometry. Thus, the fusion protein represents a readily obtainable source of biologically active dsBAFF that may prove useful in further studies on duck BAFF and its receptors.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1007/s10529-007-9543-y | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Length-Dependent Modulation of B Cell Activating Factor Transcripts in Chicken Macrophage by Viral Double-Stranded RNA.
Vaccines (Basel)
October 2023
Department of Pathobiology, College of Veterinary Medicine, Tuskegee University, Tuskegee, AL 36830, USA.
Viral double-stranded RNA (dsRNA) interacts with Retinoic-acid-inducible-gene-1 (RIG-1)-like receptors (RLRs) to induce type 1 interferons. Melanoma-derived-antigen-5 (MDA-5), an RLR, but not RIG-1, is found in chickens. Ducks express both RIG-1 and MDA-5, a possible cause of differences in susceptibility to influenza virus infection between chickens and ducks.
View Article and Find Full Text PDFMolecular cloning, expression and bioactivity of B cell activating factor (BAFF) in African ostrich.
Int Immunopharmacol
September 2015
College of Animal Science and Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, Hubei, PR China. Electronic address:
B cell activating factor (BAFF), which belongs to the tumor necrosis factor (TNF) family, is testified to play a critical role in B cell survival, proliferation, maturation and immunoglobulin secretion. In the present study, the cDNA of open reading frame (ORF) in African ostrich (Struthio camelus) BAFF (designated OsBAFF) was cloned by reverse transcription-PCR (RT-PCR). The OsBAFF gene encodes a 288-amino acid protein containing a predicted transmembrane domain and a putative furin protease cleavage site like BAFFs from chicken (cBAFF), quail (qBAFF), duck (dBAFF), goose (gBAFF) and dove (doBAFF).
View Article and Find Full Text PDFExamination of candidate genes for hypoadrenocorticism in Nova Scotia Duck Tolling Retrievers.
Vet J
February 2011
Department of Animal Science, University of California, Davis, CA 95616, USA.
Inherited hypoadrenocorticism occurs in some dog breeds including the Nova Scotia Duck Tolling Retriever (NSDTR) and is thought to be due to an immune attack on the adrenal glands. The genetic cause of this disorder in dogs has not been identified; however, many genes have been associated with hypoadrenocorticism and other immune-mediated conditions in humans including AIRE, BAFF, Casp10, CD28, CTLA-4, FASL, PTPN22, and TNFRSF6B. Microsatellite marker loci were analysed for linkage with the disease phenotype in a pedigree of NSDTRs and excluded all genes examined, the exception being CTLA-4, which was neither excluded nor shown to be associated by this analysis.
View Article and Find Full Text PDFMolecular cloning, in vitro expression and bioactivity of quail BAFF.
Vet Immunol Immunopathol
July 2009
Jiangsu Key Laboratory for Bioresource Technology, College of Life Sciences, Nanjing Normal University, Nanjing 210046, PR China.
B cell activating factor (BAFF), belonging to the TNF (tumor necrosis factor) family, is critical for B cell survival and maturation. In the present study, a quail BAFF cDNA, named qBAFF, was amplified from quail spleen by RT-PCR and RACE (rapid amplification of cDNA ends) strategies. The open reading frame (ORF) of qBAFF cDNA encodes a protein consisting of 288-amino acid.
View Article and Find Full Text PDFConstruction of bifunctional fusion proteins consisting of duck BAFF and EGFP.
Biotechnol Lett
February 2008
Jiangsu Province Key Laboratory for Molecular and Medical Biotechnology, Life Sciences College, Nanjing Normal University, Nanjing 210097, China.
We constructed fusion proteins consisting of fluorescence-enhanced green fluorescent protein (EGFP) and soluble domain of duck B-cell-activating factor of the TNF family (dsBAFF). The soluble EGFP/dsBAFF was efficiently expressed in Escherichia coli BL 21 (DE3) and was purified in milligram amounts using metal chellate affinity chromatography. The fusion protein exhibited similar fluorescence spectra with free EGFP and promoted the survival of duck bursal B cells in vitro as well as dsBAFF.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!