数字农科院2.0

Knockdown Of Bmtcp-1 Beta Delays B.mnpv Infection In Vitro

文献类型： 外文期刊

作者： Wang, Xue-yang;Shao, Zuo-min;Chen, Qian-ying;Sun, Xia;Sun, Xia;Wang, Xue-yang;Wu, Yang-chun;Xu, Zhen-ping;Xu, Zhen-ping;Wu, Yang-chun;Li, Mu-wang;Li, Mu-wang;Xu, Jia-ping

作者机构：

关键词： Bombyx mori; BmNPV; chaperonin containing t-complex polypeptide 1 beta; protein folding; response mechanism

期刊名称： FRONTIERS IN MICROBIOLOGY

ISSN： 1664-302X

年卷期： 2019 年 10 卷

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摘要： The molecular mechanism of silkworm resistance to Bombyx mori nucleopolyhedrovirus (BmNPV) infection remains unclear. The chaperonin containing t-complex polypeptide 1 (TCP-1) is essential for the folding of tubulin and actin to produce stable and functional competent protein conformation. However, little is known about this protein in silkworm. In the present study, a gene encoding the TCP-1 beta protein in silkworm was characterized, which has an open reading fragment of 1,611 bp encoding a predicted 536 amino acid residue-protein with a molecular weight of approximately 57.6 kDa containing a Cpn60_TCP1 functional domain. The sequence conservation is 81.52%. The highest level of BmTCP-1 beta mRNA expression was found in the midgut, while the lowest was in the hemolymph. To further study the function of BmTCP-1 beta, expression was knocked down with siRNA in vitro, resulting in significant downregulation of the selected cytoskeletal-related genes, actin and tubulin, which was also confirmed by overexpression of BmTCP-1 beta in BmN cells using the pIZT/V5-His-mCherry insect vector. Moreover, knockdown of BmTCP-1 beta significantly prolonged the infection process of BmNPV in BmN cells, which was also verified by overexpression of BmTCP-1 beta in BmN cells. Based on the results of the present study, we concluded that BmTCP-1 beta plays a vital role in BmNPV infection by regulating the expression of tubulin and actin. Taken together, our work provides valuable data for the clarification of the molecular mechanism of silkworm resistance to BmNPV infection.

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