Set of module vectors for stable or transient expression of heterologous genes in plants

Vyacheslavova, A. O.; Mustafaev, O. N.; Tyrin, A. A.; Shimshilashvili, Kh. R.; Berdichevets, I. N.; Shayakhmetova, D. M.; Goldenkov, M. A.; Fadeev, V. S.; Sheludko, Y. V.; Goldenkova-Pavlova, I. V.

doi:10.1134/s1022795412090098

Cited by 12 publications

(24 citation statements)

References 26 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Currently, plant cells are an attractive alternative expression system for recombinant proteins with medical purposes and are used in many leading biotechnological laboratories and companies [ 29 – 31 ]. Recent advances in this field have enabled a significant increase in the expression level of recombinant proteins [ 32 – 34 ], improvement in posttranslational modifications that fit more closely with mammalian cells [ 35 , 36 ], and approaches to directed modification of the plant genome [ 37 , 38 ]. It has become evident that plant systems possess a high potential competitive ability compared with other expression systems and are of interest for investment companies.…”

Section: Discussionmentioning

confidence: 99%

Transgenic Carrot Expressing Fusion Protein ComprisingM. tuberculosisAntigens Induces Immune Response in Mice

Permyakova

Zagorskaya

Belavin

et al. 2015

BioMed Research International

View full text Add to dashboard Cite

Tuberculosis remains one of the major infectious diseases, which continues to pose a major global health problem. Transgenic plants may serve as bioreactors to produce heterologous proteins including antibodies, antigens, and hormones. In the present study, a genetic construct has been designed that comprises the Mycobacterium tuberculosis genes cfp10, esat6 and dIFN gene, which encode deltaferon, a recombinant analog of the human γ-interferon designed for expression in plant tissues. This construct was transferred to the carrot (Daucus carota L.) genome by Agrobacterium-mediated transformation. This study demonstrates that the fusion protein CFP10-ESAT6-dIFN is synthesized in the transgenic carrot storage roots. The protein is able to induce both humoral and cell-mediated immune responses in laboratory animals (mice) when administered either orally or by injection. It should be emphasized that M. tuberculosis antigens contained in the fusion protein have no cytotoxic effect on peripheral blood mononuclear cells.

show abstract

Section: Discussionmentioning

confidence: 99%

Transgenic Carrot Expressing Fusion Protein ComprisingM. tuberculosisAntigens Induces Immune Response in Mice

Permyakova

Zagorskaya

Belavin

et al. 2015

BioMed Research International

View full text Add to dashboard Cite

show abstract

“…Plant expression vectors, carrying thermostable lichenase reporter gene fused at the 5'-terminal region to synthesized 5'-UTR sequences, were prepared in several stages. Initially, the sequence of thermostable lichenase reporter gene was obtained by PCR using pQE-licBM3 plasmid as a template [22] and forward and reverse primers with introduced BamHI and SmaI restriction sites. Primers and their sequences are shown in Table 1.…”

Section: Methodsmentioning

confidence: 99%

“…Primers and their sequences are shown in Table 1. The amplicon was cloned as a BamHI-SmaI fragment into previously obtained pPGG vector [22] with the formation of an intermediate pPGG-LicB vector.…”

Section: Methodsmentioning

confidence: 99%

“…The sequence of nucleotides with a high content of CpG dinucleotides (designated as CG) and the leader sequence of the omega (Ω) tobacco mosaic virus (TMV) were synthesized from oligonucleotides using ApaI-XhoI fragments of pPGG-LicB, pPGG-CG-LicB, and pPGG-Ω-LicB were cloned into pVIG-T vector [22], prehydrolysed with restriction endonucleases ApaI and XhoI with the formation of plant expression vectors pVIG-T-LicB, pVIG-T-CG-LicB, and pVIG-T-Ω-LicB (Fig. 1).…”

Section: Methodsmentioning

confidence: 99%

“…The amplification products of the sequences were cloned as SacI-SmaI fragments into pPGG-Ω-LicB vector, prehydrolysed with restriction endonucleases SacI and SmaI with the formation of an intermediate pPGG-Ω-CG-LicB vector, wherein the 5'-UTR Ω TMV sequence was localized between 35S RNA CaMV promoter and thermostable lichenase reporter gene, fused to the 5'-region with CG sequence. ApaI-XhoI fragment of pPGG-Ω-CG-LicB vector was cloned into pVIG-T vector [22], prehydrolysed with restriction endonucleases ApaI and XhoI with the formation of a plant expression vector pVIG-T-Ω-CG-LicB (Fig. 1).…”

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

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

RETRACTED ARTICLE: Homo and heterologous expression of the HpPKS2 gene in Hypericum perforatum and Bacopa monnieri

Khan

Verma

Parasharami

2020

Plant Cell Tiss Organ Cult

View full text Add to dashboard Cite

Set of module vectors for stable or transient expression of heterologous genes in plants

Cited by 12 publications

References 26 publications

Transgenic Carrot Expressing Fusion Protein ComprisingM. tuberculosisAntigens Induces Immune Response in Mice

Transgenic Carrot Expressing Fusion Protein ComprisingM. tuberculosisAntigens Induces Immune Response in Mice

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

RETRACTED ARTICLE: Homo and heterologous expression of the HpPKS2 gene in Hypericum perforatum and Bacopa monnieri

Contact Info

Product

Resources

About