Cloning, expression and bioactivity of duck BAFF.
B cell activating factor (BAFF) belonging to the TNF family is critical for B cell survival and maturation. In the present study, we identified a duck BAFF cDNA, named dBAFF, by RT-PCR and RACE strategies. The open reading frame (ORF) of this cDNA encodes a 288-amino acid protein containing a predicted transmembrane domain and a putative furin protease cleavage site like chicken BAFF (cBAFF), human BAFF (hBAFF) and mouse BAFF (mBAFF). The amino acid identity between biologically soluble dBAFF and cBAFF, hBAFF or mBAFF is 97, 78 and 71%, respectively. RT-PCR analysis showed the dBAFF gene is strongly expressed in the bursa of fabricius. Recombinant soluble dBAFF (dsBAFF) fused with NusA.tag was efficiently produced in Origami B (DE3) pLysS expression host strain. In vitro, purified dsBAFF was not only able to promote survival of bursa B cells, but also able to co-stimulate proliferation of mammalian B cells with anti-IgM. Furthermore, recombinant hsBAFF has a positive effect on duck bursa B cells survival. These findings indicate dBAFF plays an important role in survival and proliferation of duck B cells and because of its high conservation in the evolution, functional cross-reactivity exists between mammalian and duck BAFF.
Guan
Zheng-Bing
ZB
Jiangsu Province Key Laboratory for Molecular and Medical Biotechnology, Life Sciences College, Nanjing Normal University, Nanjing 210097, China.
Ye
Ji-Lin
JL
Dan
Wen-Bing
WB
Yao
Wen-Juan
WJ
Zhang
Shuang-Quan
SQ
D000328:Adult; D000595:Amino Acid Sequence; D000818:Animals; D053264:B-Cell Activating Factor / Q000235:genetics* / Q000502:physiology; D001483:Base Sequence; D002478:Cells, Cultured; D003001:Cloning, Molecular; D004372:Ducks / Q000235:genetics* / Q000502:physiology; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data
D053264:B-Cell Activating Factor