关键词: 蚕豆, 三叶草黄脉病毒, 核内涵体a蛋白酶, 亚细胞定位

Abstract: Clover yellow vein virus（ClYVV）is an important member of the genus of Potyvirus. As Nuclear Inclusion a Protease（NIa-Pro）is one of the most important viral-encoded proteases，it also plays an essential role in the process of most functional mature proteins generation during virus and host interaction. In this study，the NIa-Pro gene of a broad bean isolate of ClYVV，collected from Jiangsu Province in 2018，was cloned and sequenced. Sequence analysis results showed that the NIa-Pro gene was 729 bp in length and encoded a 24.3 kD protein. To clarify the subcellular localization of NIa-Pro encoded by ClYVV，we transiently expressed NIa-Pro fused to YFP under the control of the 35S promoter（YFP-NIa-Pro）in the Nicotiana benthamiana leaves by Agrobacterium infiltration. The results of laser scanning confocal microscope showed that strong fluorescence signals were observed in the cytoplasm and nucleus of epidermal cells of N. benthamiana leaves. The expression of NIa-Pro（YFP-NIa-Pro）in N. benthamiana leaves was further confirmed by Western-blot. These results showed that the NIa-Pro encoded by ClYVV localized in cytoplasm and nucleus.

Key words: broad bean, Clover yellow vein virus, Nuclear Inclusion a Protease, subcellular localization