NDLI: Comparison of effectiveness of 5′-regulatory sequences in transplastomic tobacco chloroplasts

Content Provider	Springer Nature Link
Author	Gerasymenko, I. M. Sheludko, Y. V. Klebavych, A. A. Rudas, V. A. Shakhovsky, A. M. Klein, T. M. Kuchuk, N. V.
Copyright Year	2016
Abstract	The development of tools which ensure the desired level of transgene expression in plastids is a prerequisite for the effective utilization of these plant organelles for the deployment of bioactive proteins. High-level accumulation of target proteins is considered as a positive feature of transplastomic plants, but excessive accumulation of foreign proteins may have deleterious effects on host plants. On the other hand, expression at low levels can result in ineffective phenotypes. We compared the effectiveness of different 5′-regulatory sequences in driving the expression of a reporter gene, β-glucuronidase (uidA), in tobacco chloroplasts. To achieve varying expression levels, we have chosen heterologous 5′-regulatory sequences which either differ significantly from their homologous counterparts or depend on specific nuclear encoded factors. The Medicago truncatula psbA promoter/5′-UTR supported the highest levels of protein accumulation, surpassing the other tested sequences by two to three orders of magnitude. The heterologous regulatory sequence of Phaseolus vulgaris rbcL gene was as efficient in tobacco chloroplasts as the corresponding homologous promoter/5′-UTR. The Arabidopsis thaliana ndhF promoter/5′-UTR supported as high reporter activity levels as the rbcL 5′-sequences, whereas the effectiveness of A. thaliana psbN promoter/5′-UTR was three fold lower. The characterized regulatory sequences can be utilized to establish transplastomic lines with desirable levels of target protein accumulation. The ability to control transgene expression should be useful for achieving appropriate levels of protein accumulation and thereby avoid their negative impacts on host plant physiology.
Starting Page	65
Ending Page	75
Page Count	11
File Format	PDF
ISSN	09628819
Journal	Transgenic Research
Volume Number	26
Issue Number	1
e-ISSN	15739368
Language	English
Publisher	Springer International Publishing
Publisher Date	2016-08-26
Publisher Place	Cham
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Plastome transformation Plastome promoters Plastome 5′-UTR Nicotiana tabacum Animal Genetics and Genomics Plant Genetics & Genomics Transgenics Biomedical Engineering/Biotechnology Genetic Engineering Molecular Medicine
Content Type	Text
Resource Type	Article
Subject	Genetics Animal Science and Zoology Biotechnology Agronomy and Crop Science

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