Expression of the truncated tissue plasminogen activator (K2S) gene in tobacco chloroplast

Abdoli-Nasab, Maryam; Jalali-Javaran, Mokhtar; Cusidó, Rosa M.; Palazón, Javier; Baghizadeh, Amin; Alizadeh, Houshang

doi:10.1007/s11033-013-2678-0

Cited by 15 publications

(8 citation statements)

References 45 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…20 Moreover, plant pathogens and many secondary metabolites do interfere with the overall purification at an industrial level. 21 In recent decades, the plant expression vectors were considered and being studies to deliver recombinant proteins into the host in the name of vaccine antigens. 22 Such plant-derived therapeutic proteins are cost-effective in production with high quality.…”

Section: Cloning Of Icea1 Gene Into Pbi121mentioning

confidence: 99%

Cloning and Expression of Helicobacter pylori ulcer Associated Gene - iceA1 in Brinjal (Solanum melongena L.)

Mehran¹,

Kumar

Haraprasad

et al. 2021

JPHI

View full text Add to dashboard Cite

Background: Plant biotechnology revolutionized the branch of plant sciences contributing to improving crop production. Helicobacter pylori was predominantly seen among all the sectors of the human population, closely stuck to stomach ulcers and cancer. Even though traditional vaccines are effective in preventing H. pylori, edible and non-edible parts of special transgenic plants were also being generated to express its antigens which could elicit an immune response after consuming. The induce by contact with epithelium (iceA1) protein of H. pylori is associated with virulence and stated as predisposing factor to stomach cancer. Objectives: This study intends to study the expression levels of iceA1 gene in the brinjal plant. Materials and Methods:The present study, magnified the iceA1 gene from H. pylori strain 26695 and transformed it into callus of brinjal (from leaf explants) through Agrobacterium tumefacians (EHA105). pBI121 vector was used to construct the plant expression vector, and the transgenic plants generated were further validated through gene amplification and protein expression studies. Results: Out of the 46 plants obtained from this study, five of them were found to be positive for the iceA1 expression as seen on validation assays like gene amplification and protein expression studies. Conclusion: The study could beneficially produce a vaccine candidate against H. pylori from transgenic brinjal. In Addition, it could provide, valid scientific data which can further be used by researchers.

show abstract

Section: Cloning Of Icea1 Gene Into Pbi121mentioning

confidence: 99%

Cloning and Expression of Helicobacter pylori ulcer Associated Gene - iceA1 in Brinjal (Solanum melongena L.)

Mehran¹,

Kumar

Haraprasad

et al. 2021

JPHI

View full text Add to dashboard Cite

show abstract

“…(2013) has shown that N . tabacum carrying the transgenic chloroplast had increased the expression of tPA up to 1% TSP 17 .…”

Section: Dicussionmentioning

confidence: 99%

“…The recombinant tPA expressed in the insect cells and Saccharomyces cerevisiae has shown the hyperglycosylation and the incorrect folding 13 , 14 . As a result of the high cost of the cell culture media and the virus and prion contamination, the mammalian cells are not an ideal system for the tPA expression 15 – 17 . The recombinant tPA has been expressed stably in both nucleus 15 , 16 , 18 and chloroplast 17 of plant Nicotiana tobacum .…”

Section: Introductionmentioning

confidence: 99%

“…As a result of the high cost of the cell culture media and the virus and prion contamination, the mammalian cells are not an ideal system for the tPA expression 15 – 17 . The recombinant tPA has been expressed stably in both nucleus 15 , 16 , 18 and chloroplast 17 of plant Nicotiana tobacum . However, the level of the expression in the transgenic plants is not promising.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

In silico and in vivo analyses of the mutated human tissue plasminogen activator (mtPA) and the antithetical effects of P19 silencing suppressor on its expression in two Nicotiana species

Amiri

Jalali-Javaran

Haddad

et al. 2018

Sci Rep

Self Cite

View full text Add to dashboard Cite

Human tissue-type plasminogen activator is one of the most important therapeutic proteins involved in the breakdown of blood clots following the stroke. A mutation was found at position 1541 bp (G514E) and the mutated form was cloned into the binary vector pTRAc-ERH. In silico analysis showed that this mutation might have no significant effect on the active site of the tissue plasminogen activator enzyme. Accordingly, zymography assay confirmed the serine protease activity of the mutated form and its derivatives. The expression of the mutated form was verified with/without co-agroinjection of the P19 gene silencing suppressor in both Nicotiana tabacum and N. benthamiana. The ELISA results showed that the concentration of the mutated form in the absence of P19 was 0.65% and 0.74% of total soluble protein versus 0.141% and 1.36% in the presence of P19 in N. benthamiana and N. tabacum, respectively. In N. tabacum, co-agroinjection of P19 had the synergistic effect and increased the mutated tissue plasminogen activator production two-fold higher. However, in N. benthamiana, the presence of P19 had the adverse effect of five-fold reduction in the concentration. Moreover, results showed that the activity of the mutated form and its derivatives was more than that of the purified commercial tissue plasminogen activator.

show abstract

“…Two years later, a similar achievement was attained in Nicotiana tabacum , which is a higher plant . Since then, many studies worldwide have demonstrated the successful production of human therapeutic proteins in the chloroplast including truncated tissue plasminogen activator (K2S) , interferone‐a , human insulin‐like growth factor‐1 , and cholera toxin B (CTB) subunit fused with human proinsulin .…”

Section: Introductionmentioning

confidence: 99%

Dicistronic expression of human proinsulin–protein A fusion in tobacco chloroplast

Yarbakht

Jalali-Javaran

Nikkhah

et al. 2014

Biotech and App Biochem

Self Cite

View full text Add to dashboard Cite

Different expression systems such as bacteria and mammalian cells have been used to produce pharmaceutical proteins. In recent years, the use of plants as bioreactors offers efficient and economical systems in recombinant protein production. Furthermore, because of the large number of plastid copies in plants, chloroplast engineering functions as an effective method to increase recombinant protein expression. Because the commercially available insulin for treatment does not contain C-peptide, which is of great importance for type 1 diabetic patients, the current study introduces the human proinsulin gene fused with protein A into the tobacco chloroplast genome using the biolistic method. To achieve homoplasmy, three rounds of selection and regeneration of transforming cells were performed on the medium that contained spectinomycin antibiotic and hormones. The PCR analysis indicated the presence of the proinsulin gene in transplastomic plants. The reverse-transcription PCR analysis confirmed the expression of the proinsulin-protein A fusion at the transcription level. Immunoblot assays of leaf-derived protein extracts confirmed that the target gene expression is up to 0.2% of the total soluble protein. Our study showed that protein A fusion is not as efficient as other reported fusions. The transplastomic plants were also confirmed for homoplasmy using Southern blot analysis.

show abstract

Expression of the truncated tissue plasminogen activator (K2S) gene in tobacco chloroplast

Cited by 15 publications

References 45 publications

Cloning and Expression of Helicobacter pylori ulcer Associated Gene - iceA1 in Brinjal (Solanum melongena L.)

Cloning and Expression of Helicobacter pylori ulcer Associated Gene - iceA1 in Brinjal (Solanum melongena L.)

In silico and in vivo analyses of the mutated human tissue plasminogen activator (mtPA) and the antithetical effects of P19 silencing suppressor on its expression in two Nicotiana species

Dicistronic expression of human proinsulin–protein A fusion in tobacco chloroplast

Contact Info

Product

Resources

About