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Abstract DEAD-box RNA helicase (RH) ubiquitously exists in most organisms and participates, if not all, steps of cellular RNA metabolism. Generally, containing 9 highly conserved motifs is an important standard to identify a DEAD-box RNA helicase. In the present study, the CDS of a rice (Oryza sativa L. japonica. cv. Nipponbare) gene OsRH37, encoding a putative DEAD-box RNA helicase, was cloned. The amino acid sequence of OsRH37 was analyzed by bioinformatics method and it was indicated that the amino acid sequence identity between OsRH37 and 3 other Arabidopsis thaliana DEAD-box RNA helicases including AtRH17, 37 and 52 was>70%, whereas the amino acid sequence identity between OsRH37 and DEAD-box RNA helicases from different animals was generally<60%. The overexpression vector and RNAi vector of OsRH37 were constructed, and by using rice genetic transformation system the overexpression and RNAi transgenic rice plants of OsRH37 were obtained. Six lines (66.7%) among 9 lines of OsRH37 overexpressed transgenic rice and 13 lines (92.9%) among 14 lines of OsRH37 RNAi transgenic rice were identified to be positive, respectively. The relative expression level of OsRH37 among wild type, overexpression lines, and RNAi lines were detected by sqRT-PCR. The results indicated that compared with the wild type (WT), the expression level of OsRH37 in overexpression lines was obviously up-regulated while that in RNAi lines was significantly down-regulated. To observe the subcellular localization pattern of OsRH37, the stop codon TAG deleted CDS was constructed into the transient expression vector pEarleyGate102, harboring a cyan fluorescence protein (CFP), by Gateway technology and the leaves of Nicotiana benthamiana agroinfiltrated with CFP-OsRH37 were observed under confocal microscope at 48 h post infiltrated. The results of subcellular localization analysis suggested that OsRH37 fused with CFP was mostly localized at nucleus, karyosome and cytoplasmic granular structures. The bioinformatics analysis, obtained transgenic rice plants and subcellular localization analysis of OsRH37 can provide foundational insights for the important roles of OsRH37 in the processes of the rice growth, development and stress responses.
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Received: 26 June 2015
Published: 05 October 2015
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