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Molecular and Physiological Characterization of Two Novel Multirepeat β-Thymosins from Silkworm Bombyx mori

机译:家蚕中两种新型多重复β-胸腺肽的分子和生理特性

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摘要

β-thymosin plays important roles in the development of the lymphatic system and the central nervous system in vertebrates. However, its role and function in invertebrates remain much less explored. Here, we firstly isolated a gene encoding β-thymosin in silkworm (Bombyx mori L.). Interestingly, this gene encodes two polypeptides, named as BmTHY1 and BmTHY2, via two different modes of RNA splicing. The recombinant proteins fused with an N-term GST tag were over-expressed in Escherichia coli (E. coli) and further purified to near homogenity to prepare mouse antibodies. The Western blot analysis showed that these proteins were expressed in various tissues and organs, as well as in different developmental stages. Amazingly, the expression of BmTHY2 was hugely increased during the pupae stage, indicating a specialized role in this period. The expression of these proteins was gradually decreased in BmN cells infected by BmNPV, suggesting they may play different roles in the virus infection. In addition, both BmTHY1 and BmTHY2 can interact with 14-3-3 of silkworm and Ubiquitin of BmNPV as shown by GST pull down and Co-IP assays, consistent with their roles in the regulation of the development of nervous system.
机译:β-胸腺肽在脊椎动物淋巴系统和中枢神经系统的发育中起重要作用。然而,它在无脊椎动物中的作用和功能仍然很少被探索。在这里,我们首先分离了家蚕(Bombyx mori L.)中编码β-胸腺肽的基因。有趣的是,该基因通过两种不同的RNA剪接模式编码两个多肽,分别称为BmTHY1和BmTHY2。与N端GST标签融合的重组蛋白在大肠杆菌(E. coli)中过表达,并进一步纯化至接近均一的水平,以制备小鼠抗体。蛋白质印迹分析表明,这些蛋白质在各种组织和器官以及不同的发育阶段表达。令人惊讶的是,在m期,BmTHY2的表达大大增加,表明在此期间有特殊作用。这些蛋白的表达在被BmNPV感染的BmN细胞中逐渐降低,表明它们可能在病毒感染中发挥不同的作用。此外,如GST下拉和Co-IP分析所示,BmTHY1和BmTHY2均可与蚕的14-3-3和BmNPV的泛素相互作用,这与它们在调节神经系统发育中的作用一致。

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