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J Biosci Bioeng. 2018 Jan;125(1):124-130. doi: 10.1016/j.jbiosc.2017.08.007. Epub 2017 Sep 11.

Arabidopsis thaliana cold-regulated 47 gene 5'-untranslated region enables stable high-level expression of transgenes.

Author information

1
Graduate School of Biological Science, Nara Institute of Science and Technology, 8916-5 Takayama, Ikoma, Nara 630-0192, Japan.
2
Graduate School of Pharmaceutical Sciences, Osaka University, 1-6 Yamadaoka, Suita, Osaka 565-0871, Japan.
3
Graduate School of Biological Science, Nara Institute of Science and Technology, 8916-5 Takayama, Ikoma, Nara 630-0192, Japan. Electronic address: kou@bs.naist.jp.

Abstract

Transgene expression is regulated through several steps, this study focuses on the mRNA translation step. The expression level of transgenes can be increased by 5'-untranslated region (5'UTR) sequences in certain genes which act as translational enhancers. On the other hand, translation in most mRNA species is repressed by growth, development, and stress events. There is a possibility that transgene mRNA is also repressed in these conditions, despite the use of a translational enhancer. Therefore, a consistently efficient translational enhancer is needed to develop a reliable transgene expression system. Herein we searched for mRNAs translated stably under different growth, development and environmental conditions using data sets of polysome fraction assays and microarray analysis. Correct 5'UTR sequences of candidate genes were determined by cap analysis of gene expression and we tested translational ability of the candidate 5'UTRs by reporter assays. We found the 5'UTR of cold-regulated 47 gene to be an effective translational enhancer, contributing to stable high-level expression under various conditions.

KEYWORDS:

5′UTR; Abiotic stress; Expression system; Growth and development; Plant molecular farming; Recombinant proteins; Transgenic; Translational enhancer; Translational repression

PMID:
28918993
DOI:
10.1016/j.jbiosc.2017.08.007
[Indexed for MEDLINE]

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