Switchgrass (<it>Panicum virgatum </it>L.) polyubiquitin gene (<it>PvUbi1 </it>and <it>PvUbi2</it>) promoters for use in plant transformation
<p>Abstract</p> <p>Background</p> <p>The ubiquitin protein is present in all eukaryotic cells and promoters from ubiquitin genes are good candidates to regulate the constitutive expression of transgenes in plants. Therefore, two switchgrass (<it>Panicum virgatum &...
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| Format: | Article |
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BMC
2011-07-01
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| Series: | BMC Biotechnology |
| Online Access: | http://www.biomedcentral.com/1472-6750/11/74 |
| _version_ | 1818826998395437056 |
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| author | LaFayette Peter R Hawkins Jennifer S Percifield Ryan J Joyce Blake L Liu Wusheng King Zachary R Mann David GJ Artelt Barbara J Burris Jason N Mazarei Mitra Bennetzen Jeffrey L Parrott Wayne A Stewart Charles N |
| author_facet | LaFayette Peter R Hawkins Jennifer S Percifield Ryan J Joyce Blake L Liu Wusheng King Zachary R Mann David GJ Artelt Barbara J Burris Jason N Mazarei Mitra Bennetzen Jeffrey L Parrott Wayne A Stewart Charles N |
| author_sort | LaFayette Peter R |
| collection | DOAJ |
| description | <p>Abstract</p> <p>Background</p> <p>The ubiquitin protein is present in all eukaryotic cells and promoters from ubiquitin genes are good candidates to regulate the constitutive expression of transgenes in plants. Therefore, two switchgrass (<it>Panicum virgatum </it>L.) ubiquitin genes (<it>PvUbi1 </it>and <it>PvUbi2</it>) were cloned and characterized. Reporter constructs were produced containing the isolated 5' upstream regulatory regions of the coding sequences (i.e. <it>PvUbi1 </it>and <it>PvUbi2 </it>promoters) fused to the <it>uidA </it>coding region (<it>GUS</it>) and tested for transient and stable expression in a variety of plant species and tissues.</p> <p>Results</p> <p><it>PvUbi1 </it>consists of 607 bp containing <it>cis</it>-acting regulatory elements, a 5' untranslated region (UTR) containing a 93 bp non-coding exon and a 1291 bp intron, and a 918 bp open reading frame (ORF) that encodes four tandem, head -to-tail ubiquitin monomer repeats followed by a 191 bp 3' UTR. <it>PvUbi2 </it>consists of 692 bp containing <it>cis</it>-acting regulatory elements, a 5' UTR containing a 97 bp non-coding exon and a 1072 bp intron, a 1146 bp ORF that encodes five tandem ubiquitin monomer repeats and a 183 bp 3' UTR. <it>PvUbi1 </it>and <it>PvUbi2 </it>were expressed in all examined switchgrass tissues as measured by qRT-PCR. Using biolistic bombardment, <it>PvUbi1 </it>and <it>PvUbi2 </it>promoters showed strong expression in switchgrass and rice callus, equaling or surpassing the expression levels of the CaMV <it>35S, 2x35S, ZmUbi1</it>, and <it>OsAct1 </it>promoters. GUS staining following stable transformation in rice demonstrated that the <it>PvUbi1 </it>and <it>PvUbi2 </it>promoters drove expression in all examined tissues. When stably transformed into tobacco (<it>Nicotiana tabacum</it>), the <it>PvUbi2+3 </it>and <it>PvUbi2+9 </it>promoter fusion variants showed expression in vascular and reproductive tissues.</p> <p>Conclusions</p> <p>The <it>PvUbi1 </it>and <it>PvUbi2 </it>promoters drive expression in switchgrass, rice and tobacco and are strong constitutive promoter candidates that will be useful in genetic transformation of monocots and dicots.</p> |
| first_indexed | 2024-12-19T00:36:34Z |
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| spelling | doaj.art-7ef70d9f21e54bd4ae7636fb3d6fe8492022-12-21T20:44:46ZengBMCBMC Biotechnology1472-67502011-07-011117410.1186/1472-6750-11-74Switchgrass (<it>Panicum virgatum </it>L.) polyubiquitin gene (<it>PvUbi1 </it>and <it>PvUbi2</it>) promoters for use in plant transformationLaFayette Peter RHawkins Jennifer SPercifield Ryan JJoyce Blake LLiu WushengKing Zachary RMann David GJArtelt Barbara JBurris Jason NMazarei MitraBennetzen Jeffrey LParrott Wayne AStewart Charles N<p>Abstract</p> <p>Background</p> <p>The ubiquitin protein is present in all eukaryotic cells and promoters from ubiquitin genes are good candidates to regulate the constitutive expression of transgenes in plants. Therefore, two switchgrass (<it>Panicum virgatum </it>L.) ubiquitin genes (<it>PvUbi1 </it>and <it>PvUbi2</it>) were cloned and characterized. Reporter constructs were produced containing the isolated 5' upstream regulatory regions of the coding sequences (i.e. <it>PvUbi1 </it>and <it>PvUbi2 </it>promoters) fused to the <it>uidA </it>coding region (<it>GUS</it>) and tested for transient and stable expression in a variety of plant species and tissues.</p> <p>Results</p> <p><it>PvUbi1 </it>consists of 607 bp containing <it>cis</it>-acting regulatory elements, a 5' untranslated region (UTR) containing a 93 bp non-coding exon and a 1291 bp intron, and a 918 bp open reading frame (ORF) that encodes four tandem, head -to-tail ubiquitin monomer repeats followed by a 191 bp 3' UTR. <it>PvUbi2 </it>consists of 692 bp containing <it>cis</it>-acting regulatory elements, a 5' UTR containing a 97 bp non-coding exon and a 1072 bp intron, a 1146 bp ORF that encodes five tandem ubiquitin monomer repeats and a 183 bp 3' UTR. <it>PvUbi1 </it>and <it>PvUbi2 </it>were expressed in all examined switchgrass tissues as measured by qRT-PCR. Using biolistic bombardment, <it>PvUbi1 </it>and <it>PvUbi2 </it>promoters showed strong expression in switchgrass and rice callus, equaling or surpassing the expression levels of the CaMV <it>35S, 2x35S, ZmUbi1</it>, and <it>OsAct1 </it>promoters. GUS staining following stable transformation in rice demonstrated that the <it>PvUbi1 </it>and <it>PvUbi2 </it>promoters drove expression in all examined tissues. When stably transformed into tobacco (<it>Nicotiana tabacum</it>), the <it>PvUbi2+3 </it>and <it>PvUbi2+9 </it>promoter fusion variants showed expression in vascular and reproductive tissues.</p> <p>Conclusions</p> <p>The <it>PvUbi1 </it>and <it>PvUbi2 </it>promoters drive expression in switchgrass, rice and tobacco and are strong constitutive promoter candidates that will be useful in genetic transformation of monocots and dicots.</p>http://www.biomedcentral.com/1472-6750/11/74 |
| spellingShingle | LaFayette Peter R Hawkins Jennifer S Percifield Ryan J Joyce Blake L Liu Wusheng King Zachary R Mann David GJ Artelt Barbara J Burris Jason N Mazarei Mitra Bennetzen Jeffrey L Parrott Wayne A Stewart Charles N Switchgrass (<it>Panicum virgatum </it>L.) polyubiquitin gene (<it>PvUbi1 </it>and <it>PvUbi2</it>) promoters for use in plant transformation BMC Biotechnology |
| title | Switchgrass (<it>Panicum virgatum </it>L.) polyubiquitin gene (<it>PvUbi1 </it>and <it>PvUbi2</it>) promoters for use in plant transformation |
| title_full | Switchgrass (<it>Panicum virgatum </it>L.) polyubiquitin gene (<it>PvUbi1 </it>and <it>PvUbi2</it>) promoters for use in plant transformation |
| title_fullStr | Switchgrass (<it>Panicum virgatum </it>L.) polyubiquitin gene (<it>PvUbi1 </it>and <it>PvUbi2</it>) promoters for use in plant transformation |
| title_full_unstemmed | Switchgrass (<it>Panicum virgatum </it>L.) polyubiquitin gene (<it>PvUbi1 </it>and <it>PvUbi2</it>) promoters for use in plant transformation |
| title_short | Switchgrass (<it>Panicum virgatum </it>L.) polyubiquitin gene (<it>PvUbi1 </it>and <it>PvUbi2</it>) promoters for use in plant transformation |
| title_sort | switchgrass it panicum virgatum it l polyubiquitin gene it pvubi1 it and it pvubi2 it promoters for use in plant transformation |
| url | http://www.biomedcentral.com/1472-6750/11/74 |
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