caa a22 4500 467893969 CHVBK 20180406152757.0 cr unu---uuuuu 170328e20060101xx s 000 0 eng 10.1007/s11103-005-3853-z doi (NATIONALLICENCE)springer-10.1007/s11103-005-3853-z Expression Enhancement of a Rice Polyubiquitin Gene Promoter [Elektronische Daten] [Elumalai Sivamani, Rongda Qu] An 808bp promoter from a rice polyubiquitin gene, rubi3, has been isolated. The rubi3 gene contained an open reading frame of 1140bp encoding a pentameric polyubiquitin arranged as five tandem, head-to-tail repeats of 76aa. The 1140bp 5′ UTR intron of the gene enhanced its promoter activity in transient expression assays by 20-fold. Translational fusion of the GUS reporter gene to the coding sequence of the ubiquitin monomer enhanced GUS enzyme activity in transient expression assays by 4.3-fold over the construct containing the original rubi3 promoter (including the 5′ UTR intron) construct. The enhancing effect residing in the ubiquitin monomer coding sequence has been narrowed down to the first 9nt coding for the first three amino acid residues of the ubiquitin protein. Mutagenesis at the third nucleotide of this 9nt sequence still maintains the enhancing effect, but leads to translation of the native GUS protein rather than a fusion protein. The resultant 5′ regulatory sequence, consisting of the rubi3 promoter, 5′ UTR exon and intron, and the mutated first 9nt coding sequence, has an activity nearly 90-fold greater than the rubi3 promoter only (without the 5′ UTR intron), and 2.2-fold greater than the maize Ubi1 gene promoter (including its 5′ UTR intron). The newly created expression vector is expected to enhance transgene expression in monocot plants. Considering the high conservation of the polyubiquitin gene structure in higher plants, the observed enhancement in gene expression may apply to 5′ regulatory sequences of other plant polyubiquitin genes. Springer, 2006 exon nationallicence gene expression nationallicence GUS nationallicence IME nationallicence intron nationallicence polyubiquitin nationallicence promoter nationallicence rice nationallicence GUS : β-glucuronidase nationallicence LUC : luciferase nationallicence MU : methylumbelliferone nationallicence MUG : 4-methyl-β- d-glucuronide nationallicence NOS : nopaline synthase nationallicence X-Gluc : 5-bromo-4-chloro-3-indolyl-β-d-glucuronic acid nationallicence Sivamani Elumalai Department of Crop Science, North Carolina State University, 27695-7620, Raleigh, NC, USA aut Qu Rongda Department of Crop Science, North Carolina State University, 27695-7620, Raleigh, NC, USA aut Plant Molecular Biology Kluwer Academic Publishers 60/2(2006-01-01), 225-239 0167-4412 60:2<225 2006 60 11103 https://doi.org/10.1007/s11103-005-3853-z text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s11103-005-3853-z text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Sivamani Elumalai Department of Crop Science, North Carolina State University, 27695-7620, Raleigh, NC, USA aut NATIONALLICENCE 700 1- Qu Rongda Department of Crop Science, North Carolina State University, 27695-7620, Raleigh, NC, USA aut NATIONALLICENCE 773 0- Plant Molecular Biology Kluwer Academic Publishers 60/2(2006-01-01), 225-239 0167-4412 60:2<225 2006 60 11103 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer