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鹅B细胞活化因子的分子克隆、体外表达及生物活性

Molecular cloning, in vitro expression and bioactivity of goose B-cell activating factor.

作者信息

Dan Wen-Bing, Guan Zheng-Bing, Zhang Chao, Li Bao-Cun, Zhang Jin, Zhang Shuang-Quan

机构信息

Jiangsu Province Key Laboratory for Molecular and Medical Biotechnology, Life Sciences College, Nanjing Normal University, Nanjing 210097, China.

出版信息

Vet Immunol Immunopathol. 2007 Jul 15;118(1-2):113-20. doi: 10.1016/j.vetimm.2007.03.012. Epub 2007 Apr 1.

Abstract

B-cell activating factor (BAFF), belonging to the TNF family, is critical for B cell survival and maturation. cDNA of goose BAFF (gBAFF) was amplified from goose spleen by RT-PCR. The open reading frame (ORF) of gBAFF encodes a protein of 288-amino acid. The gBAFF shows 98, 92, 44 and 55% amino acid sequence identity with duck (dBAFF), chicken (cBAFF), mouse (mBAFF) and human BAFF (hBAFF), respectively. RT-PCR results showed that gBAFF mRNA is expressed in thymus and more highly expressed in the bursa of Fabricius and spleen. Recombinant soluble gBAFF (gsBAFF) expressed in Escherichia coli has molecular weight of approximately 19kDa. In vitro, purified gsBAFF was able to promote bursa B cells survival/proliferation in goose, duck and chicken. Furthermore, recombinant dsBAFF and csBAFF have a positive effect on goose, duck and chicken bursa B cells survival/proliferation. These findings indicate that gBAFF plays an important role in the survival/proliferation of goose B cells and, owing to its high evolutionary conservation, functional cross-reactivity exists between chicken, duck and goose BAFF.

摘要

B细胞活化因子(BAFF)属于肿瘤坏死因子家族,对B细胞的存活和成熟至关重要。通过逆转录聚合酶链反应(RT-PCR)从鹅脾脏中扩增出鹅BAFF(gBAFF)的互补DNA(cDNA)。gBAFF的开放阅读框(ORF)编码一个288个氨基酸的蛋白质。gBAFF与鸭(dBAFF)、鸡(cBAFF)、小鼠(mBAFF)和人BAFF(hBAFF)的氨基酸序列同一性分别为98%、92%、44%和55%。RT-PCR结果显示,gBAFF mRNA在胸腺中表达,在法氏囊和脾脏中表达更高。在大肠杆菌中表达的重组可溶性gBAFF(gsBAFF)的分子量约为19kDa。在体外,纯化的gsBAFF能够促进鹅、鸭和鸡的法氏囊B细胞存活/增殖。此外,重组dBAFF和cBAFF对鹅、鸭和鸡的法氏囊B细胞存活/增殖有积极作用。这些发现表明,gBAFF在鹅B细胞的存活/增殖中起重要作用,并且由于其高度的进化保守性,鸡、鸭和鹅的BAFF之间存在功能交叉反应。

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