title: Molecular cloning, expression and bioactivity of B cell activating factor (BAFF) in African ostrich.
abstract: 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). RT-PCR analysis showed that the OsBAFF gene is strongly expressed in the bursa of Fabricius, thymus, spleen, and bone marrow. The soluble OsBAFF had been cloned into pET28a. SDS-PAGE and Western blotting analysis confirmed that the soluble fusion protein His-OsBAFF was efficiently expressed in Escherichia coli Rosset (DE3). In vitro, purified OsBAFF was not only able to promote the survival of African ostrich bursal lymphocytes, but also able to co-stimulate proliferation of mouse splenic B cells. The expression of OsBAFF in lymphocyte cells was higher than the control after LPS stimulation. These findings indicated that OsBAFF plays an important role in survival and proliferation of African ostrich bursal lymphocytes, which may provide valuable information for research into the immune system of African ostrich and OsBAFF may serve as a potential immunologic factor for enhancing immunological efficacy in African ostrich and any other birds.
authors.affiliation: College of Animal Science and Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, Hubei, PR China; Hubei Key Laboratory of Animal Embryo and Molecular Breeding, Institute of Animal Husbandry and Veterinary, Hubei Academy of Agricultural Sciences, Wuhan 430064, Hubei, PR China.
authors.forename: Keli
authors.initials: K
authors.lastname: Yang
authors.affiliation: College of Animal Science and Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, Hubei, PR China.
authors.forename: Ke
authors.initials: K
authors.lastname: Xiao
authors.affiliation: College of Animal Science and Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, Hubei, PR China.
authors.forename: Haibo
authors.initials: H
authors.lastname: Huang
authors.affiliation: College of Animal Science and Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, Hubei, PR China.
authors.forename: Shun
authors.initials: S
authors.lastname: Lu
authors.affiliation: College of Animal Science and Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, Hubei, PR China; College of Veterinary Medicine, Auburn University, Auburn 36849, AL, USA.
authors.forename: Juming
authors.initials: J
authors.lastname: Zhong
authors.affiliation: College of Animal Science and Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, Hubei, PR China.
authors.forename: Abdur Rahman
authors.initials: AR
authors.lastname: Ansari
authors.affiliation: College of Animal Science and Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, Hubei, PR China.
authors.forename: Haseeb
authors.initials: H
authors.lastname: Khaliq
authors.affiliation: College of Animal Science and Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, Hubei, PR China.
authors.forename: Hui
authors.initials: H
authors.lastname: Song
authors.affiliation: College of Animal Science and Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, Hubei, PR China.
authors.forename: Huazhen
authors.initials: H
authors.lastname: Liu
authors.affiliation: College of Animal Science and Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, Hubei, PR China. Electronic address: pengkm@mail.hzau.edu.cn.
authors.forename: Kemei
authors.initials: K
authors.lastname: Peng
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D053264:B-Cell Activating Factor / Q000235:genetics* / Q000276:immunology / Q000494:pharmacology; D001402:B-Lymphocytes / Q000276:immunology / Q000378:metabolism; D001483:Base Sequence; D015153:Blotting, Western; D002060:Bursa of Fabricius / Q000276:immunology; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000276:immunology; D002478:Cells, Cultured; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D005786:Gene Expression Regulation; D008070:Lipopolysaccharides / Q000494:pharmacology; D051379:Mice; D008969:Molecular Sequence Data; D016366:Open Reading Frames / Q000235:genetics; D009928:Organ Specificity; D010802:Phylogeny; D060888:Real-Time Polymerase Chain Reaction; D020310:Struthioniformes / Q000235:genetics* / Q000276:immunology*
chemical_list: D053264:B-Cell Activating Factor; D018076:DNA, Complementary; D008070:Lipopolysaccharides; C433788:lipopolysaccharide, E. coli O26-B6
keywords: B cell survival; BAFF; Expression; Ostrich (Struthio camelus)