Development of dual reporter vector system for estimating translational activity of regulatory elements

For the needs of modern biotechnology, a quantitative approach to the control of regulatory elements at all stages of gene expression has long become indispensable. Such a control regime is impossible without a quantitative analysis of the role of each regulatory element or pattern used. Therefore,...

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Bibliographic Details
Published in:	BMC plant biology Vol. 22; no. 1; pp. 1 - 356
Main Authors:	Suhorukova, Aleksandra V, Tyurin, Alexander A, Pavlenko, Olga S, Mustafayev, Orkhan N, Sinelnikov, Igor G, Goldenkova-Pavlova, Irina V
Format:	Journal Article
Language:	English
Published:	London BioMed Central Ltd 21-07-2022 BioMed Central BMC
Subjects:	Biotechnology Botanical research Cell cycle E coli Enhancers Enzymatic activity Enzymes Experiments Gene expression Genes Genetic vectors High throughput screening Lichenase Plant genetics Proteins Quantitative analysis Regulatory elements Regulatory sequences Transcription Transient expression Translation Vector β-glucuronidase Russia
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Summary:	For the needs of modern biotechnology, a quantitative approach to the control of regulatory elements at all stages of gene expression has long become indispensable. Such a control regime is impossible without a quantitative analysis of the role of each regulatory element or pattern used. Therefore, it seems important to modify and develop the accuracy, reproducibility, and availability of methods for quantifying the contribution of each regulatory code to the implementation of genetic information. A new vector system for transient expression in plants is described; this system is intended for quantitative analysis of the contribution of regulatory elements to transcription and translation efficiencies. The proposed vector comprises two expression cassettes carrying reporter genes (of the Clostridium thermocellum thermostable lichenase and E. coli[beta]-glucuronidase) under the control of different promoters. Herewith we also propose a new method for quantification of the effect of tested regulatory elements on expression, which relies on assessment of the enzyme activities of reporter proteins taking into account the transcription of their genes. In our view, this approach makes it possible to precisely determine the amounts of reporter proteins and their transcripts at all stages of expression. The efficiency of the proposed system has been validated by the analysis of the roles of known translation enhancers at the stages of transcription and translation.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	1471-2229 1471-2229
DOI:	10.1186/s12870-022-03735-1