Inducible expression of human papillomavirus‐16 L1 capsomeres in the plastomes of <i>Nicotiana tabacum</i>: Transplastomic plants develop normal flowers and pollen

Latif, Sara; Gottschamel, Johanna; Syed, Tahira; Younus, Iqra; Gull, Kehkshan; Sameeullah, Muhammad; Batool, Neelam; Lössl, Andreas Günter; Mariz, Filipe Colaço; Müller, Martin; Mirza, Bushra; Waheed, Mohammad Tahir

doi:10.1002/bab.2136

Cited by 6 publications

(5 citation statements)

References 44 publications

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“…End-to-end PCR for detecting whether all plastomes are in transformed state. The method has been used previously for investigating that all plastid genomes are in transformed state and no wild-type plastid genome is left [ 90 , 91 ]. For this purpose, a pair of primers was used which gave positive results for both wild-type and transgenic lines with different amplicon sizes.…”

Section: Methodsmentioning

confidence: 99%

Plastidial Expression of 3β-Hydroxysteroid Dehydrogenase and Progesterone 5β-Reductase Genes Confer Enhanced Salt Tolerance in Tobacco

Sameeullah

Yıldırım

Aslam

et al. 2021

IJMS

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The short-chain dehydrogenase/reductase (SDR) gene family is widely distributed in all kingdoms of life. The SDR genes, 3β-hydroxysteroid dehydrogenase (3β-HSD) and progesterone 5-β-reductases (P5βR1, P5βR2) play a crucial role in cardenolide biosynthesis pathway in the Digitalis species. However, their role in plant stress, especially in salinity stress management, remains unexplored. In the present study, transplastomic tobacco plants were developed by inserting the 3β-HSD, P5βR1 and P5βR2 genes. The integration of transgenes in plastomes, copy number and transgene expression at transcript and protein level in transplastomic plants were confirmed by PCR, end-to-end PCR, qRT-PCR and Western blot analysis, respectively. Subcellular localization analysis showed that 3β-HSD and P5βR1 are cytoplasmic, and P5βR2 is tonoplast-localized. Transplastomic lines showed enhanced growth in terms of biomass and chlorophyll content compared to wild type (WT) under 300 mM salt stress. Under salt stress, transplastomic lines remained greener without negative impact on shoot or root growth compared to the WT. The salt-tolerant transplastomic lines exhibited enhanced levels of a series of metabolites (sucrose, glutamate, glutamine and proline) under control and NaCl stress. Furthermore, a lower Na+/K+ ratio in transplastomic lines was also observed. The salt tolerance, mediated by plastidial expression of the 3β-HSD, P5βR1 and P5βR2 genes, could be due to the involvement in the upregulation of nitrogen assimilation, osmolytes as well as lower Na+/K+ ratio. Taken together, the plastid-based expression of the SDR genes leading to enhanced salt tolerance, which opens a window for developing saline-tolerant plants via plastid genetic engineering.

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Section: Methodsmentioning

confidence: 99%

Plastidial Expression of 3β-Hydroxysteroid Dehydrogenase and Progesterone 5β-Reductase Genes Confer Enhanced Salt Tolerance in Tobacco

Sameeullah

Yıldırım

Aslam

et al. 2021

IJMS

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“…A probe of 795 bp (Gene G418) was used and was purified with a PCR purification kit. The probe was labelled and hybridized using the DIG High Prime DNA Labelling and Detection Starter kit II for chemiluminescent detection with NBT/BCIP (Roche, Mannheim, Germany), following the manufacturer's instructions [31].…”

Section: Southern Blotting Analysismentioning

confidence: 99%

Trichoderma harzianum Cellulase Gene thph2 Affects Trichoderma Root Colonization and Induces Resistance to Southern Leaf Blight in Maize

Lang,

Chen

2023

JoF

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Trichoderma, widely distributed all over the world, is commonly found in soil and root ecosystems. It is a group comprising beneficial fungi that improve plant disease resistance and promote plant growth. Studies have shown that Trichoderma cellulases can also improve plant disease resistance. Based on previous studies, we reported that a C6 zinc finger protein (Thc6) regulates two cellulase genes, thph1 and thph2, to induce ISR responses in plants. Therefore, in this study, we focused on the role of thph2 in the colonization of maize roots by T. harzianum and the induction of systemic resistance against southern leaf blight. The results showed that thph2 had a positive regulatory effect on the Trichoderma colonization of maize roots. After the root was treated with Trichoderma, the leaf defense genes AOS, LOX5, HPL, and OPR1 were expressed to resist the attack of Cochliobolus heterostrophus. The pure Thph2 protein also resulted in a similar induction activity of the AOS, LOX5, HPL, and OPR1 expression in maize roots, further demonstrating that thph2 can induce plant defense responses and that signal transduction occurs mainly through the JA signaling pathway.

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“…[ 100 ] With this ethanol‐inducted T7 system, several pantheon antigens were also produced by tobacco seedling, such as dengue virus EDIII, [ 101 ] tuberculosis ESAT‐6 antigen, [ 102 ] and human papillomavirus‐16 L1 capsomeres. [ 103 ] In addition to the abovementioned inducible T7 system, a theophylline‐reponsive riboswitch T7 system, named as RNA amplification‐enhanced riboswitch (RAmpER), was constructed in tobacco. What's more, the RAmpER was characterized with the nuclear independence, that is, the T7 RNAP and T7 promoter‐driven target gene was all integrated into chloroplast genome, which could avoid the problems of nuclear transgene, especially the issue of transgenetic pollen transmission in environment.…”

Section: Plantmentioning

confidence: 99%

Construction and Application of Orthogonal T7 Expression System in Eukaryote: An Overview

Wang

Yan

et al. 2022

Advanced Biology

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to its high efficiency and yield for protein synthesis. [12] Among the commonlyused protein expression systems in E. coli (Table 1), T7 expession system prepared more than 90% of proteins in protein data bank (PDB) due to its high yield and translation efficiency for target protein. [13] The T7 system had been applied extensively for target protein and mRNA synthesis in prokaryote, eukaryote, and cell free system. [2] UP to now, it was also becoming an enabling tool for on-going synthetic biology, metabolic engineering, biomedicine, enzyme engineering, and so on. [14][15][16][17] Although T7 expression system had been developed successfully in various prokaryotes, [2] the construction of T7 expression system in eukaryote was still on-going, which attributed to the innate feature of transcription-translation system in eukaryote, for example, in prokaryote, the transcription was coupled with translation, while in eukaryote, mRNA was synthesized and modified in nucleus, then exporting to cytoplasm for protein synthesis. Thus far, there was no systematic and complete review about T7 expression system construction in eukaryote. In the present paper, the development of T7 expression system in eukaryote was reviewed, including its construction in animal (mammalian cells, trypanosomatid protozoa, Xenopus oocytes, zebrafish), plant, and microorganism and its application in vaccine preparation and gene therapy. We also discussed the innate challenges of T7 expression system construction in eukaryote and its recent development in vaccine production and gene therapy. Animal Mammalian CellsDue to the pivotal role in life science research, clinical research, gene therapy, and vaccine production, mammalian cells were first selected as the eukaryotic host to construct T7 expression system; so far, mammalian cells have experienced a deep and systematic study on the construction of T7 expression system. In 1986, a vaccinia/T7 hybrid virus expression system (VT7 expression system) was constructed in CV-1 monkey kidney cells, in which two expression vectors co-infected host cells, including a recombinant vaccinia virus containing vaccinia promoter-driven T7 RNAP and a plasmid containing T7 promoter-driven target gene (Figure 1A). [26] Vaccinia is a cytoplasmic DNA virus, that is, the genome DNA of vaccinia exists in host cytoplasm. Vaccinia genome encodes all replication andThe T7 system is an orthogonal transcription-system, which is characterized by simplicity, higher efficiency, and higher processivity, and it is used for protein or mRNA synthesis in various biological-systems. In comparison with prokaryotes, the construction of the T7 expression system is still ongoing in eukaryotes, but it shows greatly applicable prospects. In the present paper, development of T7 expression system construction in eukaryotes is reviewed, including its construction in animal (mammalian cells, trypanosomatid protozoa, Xenopus oocytes, zebrafish), plant, and microorganism and its application in vaccine production and gene therapy. In addition, the...

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Inducible expression of human papillomavirus‐16 L1 capsomeres in the plastomes of Nicotiana tabacum: Transplastomic plants develop normal flowers and pollen

Cited by 6 publications

References 44 publications

Plastidial Expression of 3β-Hydroxysteroid Dehydrogenase and Progesterone 5β-Reductase Genes Confer Enhanced Salt Tolerance in Tobacco

Plastidial Expression of 3β-Hydroxysteroid Dehydrogenase and Progesterone 5β-Reductase Genes Confer Enhanced Salt Tolerance in Tobacco

Trichoderma harzianum Cellulase Gene thph2 Affects Trichoderma Root Colonization and Induces Resistance to Southern Leaf Blight in Maize

Construction and Application of Orthogonal T7 Expression System in Eukaryote: An Overview

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