Motif Analysis and Subcellular Localization of OsUF in Oryza sativa
DING Zuo-Mei, WANG Yuan-Yuan, SHI Chao-Nan, WANG Hong, ZHANG Cheng-Long, ZHENG Xiao-Yuan, ZHANG Chao, WU Zu-Jian, WU Jian-Guo*
Fujian Province Key Laboratory of Plant Virology/Institute of Plant Virology/Vector-borne Virus Research Center, Fujian Agriculture and Forestry University, Fuzhou 350002, China
Abstract：E3 ubiquitin ligases, a set of crucial component of ubiquitination pathway, are involved in many aspects of biological process. In order to seek more genes which have key function in antiviral response, this study based on the previous research achievements amplified the Oryza sativa ubiquitin fusion protein (OsUF) cDNA sequence (GenBank No. XM_015757177.2) by using reverse transcription-PCR (RT-PCR) method. The bioinformatics analysis showed that OsUF contained 5 ubiquitin motifs (Ubiquitin, Rad60-SLD, Ubiquitin-2, Rad60-SLD-2 and DUF2407), and 2 non-ubiquitin motif (Ribosomal_L4F01020 and zf-C4pol). Then the full-length of OsUF cDNA was reconstituted into pEarleyGate103, a transient expression vector fused expression with green fluorescent protein (GFP). Agrobacterium-mediated transient expression of OsUF was performed in the epidermal cells of Nicotiana benthamina leaves. Subcellular localization and 4',6-diamidino-2-phenylindole (DAPI) dyeing showed that OsUF protein was localized in the nucleus. Western blotting showed that the GFP-fused OsUF protein was expressed. This analysis of motif and subcellular localization pattern of OsUF may provide key insights into the elucidation of biological function of OsUF and gives more information to learn the mechanism of antiviral response in rice.
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