Characterization of Nicotiana tabacum plants expressing hybrid genes of cyanobacterial Δ9 or Δ12 acyl-lipid desaturases and thermostable lichenase

Gerasymenko, I. M.; Sakhno, L. O.; Kyrpa, T. N.; Остапчук, А. М.; Hadjiev, T. A.; Goldenkova-Pavlova, I. V.; Sheludko, Y. V.

doi:10.1134/s1021443715030073

Cited by 12 publications

(14 citation statements)

References 27 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…The native (desC) and hybrid (desC-egfp) genes in the pVIG-Lch-D9 and pVIG-Lch-D9E vectors, respectively, had a well-characterized signal in the 5' region ensuring the localization of the target proteins into chloroplasts ( Fig. 1) (Gerasymenko et al 2015). In the pVIG-LeB4-D9-ER and pVIG-LeB4-D9E-ER expression vectors, a sequence corresponding to the LeB4 signal peptide was cloned into the 5' regions of the target genes (desC and desC-egfp) for localizing the protein product to the ER, and the fragment encoding the signal of protein retention in the ER was introduced into the 3' region ( Fig.…”

Section: Resultsmentioning

confidence: 99%

“…Gene engineering approaches, such as constitutive expression of heterologous desaturase genes (Peng et al 2018, Shi et al 2018 and the RNA interference technology as a means to inhibit the expression of the target desaturase gene (Wang et al 2010), are often used as convenient models to study the role of desaturases, including Δ9 desaturase, in plant physiology. Specifically, it has been convincingly demonstrated that constitutive expression of heterologous Δ9 desaturase genes in transgenic plants leads to effective conversion of stearic acid (18:0) into oleic acid (18:1), which is accompanied by considerable changes in the unsaturation of FAs, including polyunsaturated FAs (PUFAs) in membrane lipids (Orlova et al 2003, Craig et al 2008, Gerasymenko et al 2015.It is currently assumed that plant enzymes of this group are characterized by diverse lipid substrates, region-specific desaturation, and the capacity for functioning in different cell compartments, which deserves further study.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Altered fatty acid composition of Nicotiana benthamiana and Nicotiana excelsior leaves under transient overexpression of the cyanobacterial desC gene

Berestovoy¹,

Pavlenko²,

Tyurin³

et al. 2020

Biologia plant.

View full text Add to dashboard Cite

Transient heterologous gene expression in two model plant species, Nicotiana benthamiana and N. excelsior, has been used to study the localization of the heterologous Δ9 acyl-lipid desaturase (Δ9 desaturase) of Synechococcus vulcanus in different cell compartments and its functional activity in the cases of the cytosol, chloroplast, and endoplasmic reticulum (ER) localization. The functional activity and substrate specificity of the heterologous desaturase under the conditions of transient expression have been confirmed by comparison of fatty acid (FA) profiles. The Δ9 desaturase, responsible for the synthesis of oleic and palmitoleic acids, has also been shown to strongly promote the accumulation of polyunsaturated FAs. The results convincingly demonstrate that the Δ9 desaturase of the thermophilic cyanobacterium transiently expressed in two Nicotiana species considerably alters lipid metabolism in their leaves towards a higher FA unsaturation. The functional activity of Δ9 desaturase depends on both the model plant species, N. benthamiana or N. excelsior, and the cellular localization of the enzyme. The method of transient expression of heterologous genes in plants is highly effective, inexpensive, and not time-consuming, which makes it attractive for estimating the functional activity and/or substrate specificity of heterologous desaturases.

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Altered fatty acid composition of Nicotiana benthamiana and Nicotiana excelsior leaves under transient overexpression of the cyanobacterial desC gene

Berestovoy¹,

Pavlenko²,

Tyurin³

et al. 2020

Biologia plant.

View full text Add to dashboard Cite

show abstract

“…This makes the expression reporter system, based on lichenase, highly specific and effective to monitor the accumulation of recombinant proteins in plants. It should also be noted that a compact spatial structure of lichenase catalytic domain allows, in many cases, to keep the activity of a target sequence that was fused to lichenase unchanged [23].…”

Section: Discussionmentioning

confidence: 99%

“…with a suspension of agrobacteria, carrying respective vectors, as described previously [23]. Plant regeneration was conducted on MS medium containing 20 g/L sucrose, 1 mg/L BAP (Sigma, United States), 0.1 mg/L NAA (Sigma), 500 mg/L cefotaxime, and 100 mg/L kanamycin sulfate as a selective agent.…”

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

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

Tyurin

Кабардаева

Gra

et al. 2016

Russ J Plant Physiol

View full text Add to dashboard Cite

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.

show abstract

Combinatorial biosynthesis of small molecules in plants: Engineering strategies and tools

Gerasymenko

Sheludko

Fräbel

et al. 2019

Methods in Enzymology

View full text Add to dashboard Cite

Characterization of Nicotiana tabacum plants expressing hybrid genes of cyanobacterial Δ9 or Δ12 acyl-lipid desaturases and thermostable lichenase

Cited by 12 publications

References 27 publications

Altered fatty acid composition of Nicotiana benthamiana and Nicotiana excelsior leaves under transient overexpression of the cyanobacterial desC gene

Altered fatty acid composition of Nicotiana benthamiana and Nicotiana excelsior leaves under transient overexpression of the cyanobacterial desC gene

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

Combinatorial biosynthesis of small molecules in plants: Engineering strategies and tools

Contact Info

Product

Resources

About