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

编号 040035801

推送时间 20220829

研究领域 森林培育 

年份 2022 

类型 期刊 

语种 英语

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

来源期刊 BMC Plant Biology

期 第358期

发表时间 20220721

关键词 High throughput screening;  Vector;  Lichenase;  β-glucuronidase;  Regulatory elements;  Transient
expression; 

摘要 Background
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.
Results
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 β-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.
Conclusions
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.

服务人员 孙小满

服务院士 尹伟伦

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