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家蚕 V-ATPase 16 kDa 亚基 c 的表达与定位。

Expression and localization of Bombyx mori V-ATPase 16 kDa subunit c.

机构信息

Institute of Life Sciences, Jiangsu University, No. 301 Xuefu Road, Zhenjiang 212013, P. R. China.

出版信息

Z Naturforsch C J Biosci. 2010 Jan-Feb;65(1-2):119-26. doi: 10.1515/znc-2010-1-219.

Abstract

V-ATPase plays a central role in lepidopteran midgut ion transport physiology, and lepidopteran midgut turned out to be a model tissue for the study of V-ATPase. In the present study, the 5'-RACE method is used to obtain the 5'-UTR of V-ATPase c subunit gene from Bombyx mori. Sequence analysis of the promoter region and 3'-UTR of V-ATPase c subunit gene revealed that the transcription of the V-ATPase c subunit gene may be regulated by multi-ways. RT-PCR analysis showed that B. mori V-ATPase c subunit mRNA expresses in the whole developmental stages of B. mori. We also constructed a transient vector to determine the subcellular localization of the B. mori V-ATPase c subunit, and the result demonstrated that it is located in the membrane and some specific regions of BmN cells. Real-time PCR analysis further indicated that the c subunit mRNA expression was upregulated significantly at 24 and 72 h in the midguts of resistant B. mori larvae after being inoculated with B. mori nucleopolyhedrovirus, suggesting that it may be related to the immune response against virus infection.

摘要

V-ATPase 在鳞翅目昆虫中发挥着核心作用,其肠道是研究 V-ATPase 的模式组织。本研究采用 5'-RACE 方法从家蚕中获得 V-ATPase c 亚基基因的 5'-UTR。对 V-ATPase c 亚基基因启动子区和 3'-UTR 的序列分析表明,V-ATPase c 亚基基因的转录可能受到多种方式的调控。RT-PCR 分析表明,家蚕 V-ATPase c 亚基 mRNA 在整个家蚕发育阶段都有表达。我们还构建了瞬时载体来确定家蚕 V-ATPase c 亚基的亚细胞定位,结果表明它位于 BmN 细胞的膜和一些特定区域。实时 PCR 分析进一步表明,在感染家蚕核型多角体病毒后,抗性家蚕幼虫中肠的 c 亚基 mRNA 表达在 24 和 72 小时显著上调,表明其可能与对病毒感染的免疫反应有关。

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