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Molecular cloning, expression and bioactivity of B cell activating factor (BAFF) in African ostrich

机译：非洲鸵鸟B细胞活化因子（BAFF）的分子克隆，表达及生物活性

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摘要

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 inununoglobulin secretion. In the present study, the cDNA of open reading frame (ORE) 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. (C) 2015 Elsevier B.V. All rights reserved.
机译：B细胞活化因子（BAFF）属于肿瘤坏死因子（TNF）家族，经证实在B细胞存活，增殖，成熟和免疫球蛋白分泌中起关键作用。在本研究中，通过逆转录PCR（RT-PCR）克隆了非洲鸵鸟（Struthio camelus）BAFF（命名为OsBAFF）的开放阅读框（ORE）的cDNA。 OsBAFF基因编码一个288个氨基酸的蛋白质，包含一个预测的跨膜结构域和一个假定的弗林蛋白酶蛋白酶切割位点，如来自鸡（cBAFF），鹌鹑（qBAFF），鸭（dBAFF），鹅（gBAFF）和鸽子（doBAFF）的BAFF。 RT-PCR分析表明，OsBAFF基因在法氏囊，胸腺，脾脏和骨髓的囊中强烈表达。可溶性OsBAFF已被克隆到pET28a中。 SDS-PAGE和蛋白质印迹分析证实可溶性融合蛋白His-OsBAFF在大肠杆菌Rosset（DE3）中有效表达。在体外，纯化的OsBAFF不仅能够促进非洲鸵鸟法氏囊淋巴细胞的存活，而且能够共同刺激小鼠脾脏B细胞的增殖。 LPS刺激后，淋巴细胞中OsBAFF的表达高于对照组。这些发现表明，OsBAFF在非洲鸵鸟法氏囊淋巴细胞的存活和增殖中起着重要作用，这可能为非洲鸵鸟免疫系统的研究提供有价值的信息，并且OsBAFF可能是增强非洲鸵鸟免疫力的潜在免疫学因子。任何其他鸟类。 （C）2015 Elsevier B.V.保留所有权利。

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