Efficient expression of a heterologous gene in plants depends on the nucleotide composition of mRNA’s 5'-region

Efficient expression of a heterologous gene in plants depends on the nucleotide composition of... The contribution of nucleotide composition of mRNA 5'-region to the efficiency of expression at transcriptional and translational levels was studied in transgenic tobacco plants (Nicotiana tabacum L., cultivar Petit Havana) using a thermostable lichenase reporter gene. Synthetic sequence that contains CG-rich motifs, typical for 5'-region of plant genes, identified in silico, was constructed. Transgenic plant lines of N. tabacum were obtained; they contain thermostable lichenase reporter gene that is under control of the constitutive 35S RNA CaMV promoter and additional regulatory element: synthetic CG-rich sequence, which functions as the 5'-UTR (untranslated region) of the reporter gene mRNA or as 5'-region of the hybrid gene coding sequence, wherein the synthetic sequence is fused with the sequence of the reporter gene in its reading frame. Results of the comparative analysis of mRNA and protein levels in the obtained lines of transgenic plants showed that the synthetic CG-rich sequence significantly increases the level of transcription of the reporter gene and appear to have no negative effect on the efficiency of reporter mRNA translation, which may be due to the peculiarities of its nucleotide composition and structure, namely, due to the presence of motifs that are specific for 5'-regions of plant genes, as well as due to the properties of the secondary structure— the absence of hairpin structures with a high energy of formation. It was experimentally confirmed for the first time that the 5'-region of the genes with a high content of CpG dinucleotides can help to increase the transcription level of genes in plants. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Russian Journal of Plant Physiology Springer Journals

Efficient expression of a heterologous gene in plants depends on the nucleotide composition of mRNA’s 5'-region

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Publisher
Pleiades Publishing
Copyright
Copyright © 2016 by Pleiades Publishing, Ltd.
Subject
Life Sciences; Plant Physiology; Plant Sciences
ISSN
1021-4437
eISSN
1608-3407
D.O.I.
10.1134/S1021443716030158
Publisher site
See Article on Publisher Site

Abstract

The contribution of nucleotide composition of mRNA 5'-region to the efficiency of expression at transcriptional and translational levels was studied in transgenic tobacco plants (Nicotiana tabacum L., cultivar Petit Havana) using a thermostable lichenase reporter gene. Synthetic sequence that contains CG-rich motifs, typical for 5'-region of plant genes, identified in silico, was constructed. Transgenic plant lines of N. tabacum were obtained; they contain thermostable lichenase reporter gene that is under control of the constitutive 35S RNA CaMV promoter and additional regulatory element: synthetic CG-rich sequence, which functions as the 5'-UTR (untranslated region) of the reporter gene mRNA or as 5'-region of the hybrid gene coding sequence, wherein the synthetic sequence is fused with the sequence of the reporter gene in its reading frame. Results of the comparative analysis of mRNA and protein levels in the obtained lines of transgenic plants showed that the synthetic CG-rich sequence significantly increases the level of transcription of the reporter gene and appear to have no negative effect on the efficiency of reporter mRNA translation, which may be due to the peculiarities of its nucleotide composition and structure, namely, due to the presence of motifs that are specific for 5'-regions of plant genes, as well as due to the properties of the secondary structure— the absence of hairpin structures with a high energy of formation. It was experimentally confirmed for the first time that the 5'-region of the genes with a high content of CpG dinucleotides can help to increase the transcription level of genes in plants.

Journal

Russian Journal of Plant PhysiologySpringer Journals

Published: Jun 24, 2016

References

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