Designing a new marker-free and tissue-specific platform for molecular farming applications View Full Text


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Article Info

DATE

2015-02-08

AUTHORS

Motahhareh Mohsenpour, Masoud Tohidfar, Nadali Babaeian Jelodar, Gholamreza Salehi Jouzani

ABSTRACT

Tissue-specific expression of recombinant proteins in transgenic plants is of particular importance. Production of tissue-specific transplastomic plants is challenging due to lack of tissue-specific promoter in the plastome. The objective of the present study was to produce a transgenic platform enhancing tissue-specific transgene expression in the green organs of Nicotiana tabacum Cv. Jafarabadi. A bacteria/plant hybrid sigma factor sequence (hsig) under control of the PEP Carboxylase promoter (PEPC) was designed and cloned in a plasmid construct without selectable marker gene and finally transformed into the nucleus genome by Agrobacterium. Transformation was confirmed by PCR, RT-PCR, southern blot and real-time PCR analysis. The resulted platform was re-transformed using a plastid construct pFNGi containing gfp (reporter gene) and neo genes (a kanamycin resistance selectable marker gene). Expression of GFP was assayed under Epifluorescence microscopy. The results showed that the constructed platform was functional, and the GFP was expressed. Finally, it can be concluded that application of the hybrid sigma factor (hsig) under control of a tissue-specific promoter could be a suitable platform for tissue-specific expression of plastome integrated transgenes, and also it is proposed to use other more safer marker genes for plastid transformation instead of the antibiotic resistance gene. More... »

PAGES

433-440

References to SciGraph publications

  • 2003-08. Marker-free transgenic plants in PLANT CELL, TISSUE AND ORGAN CULTURE (PCTOC)
  • 2006-11-14. Disruption of essential plastid gene expression caused by T7 RNA polymerase-mediated transcription of plastid transgenes during early seedling development in TRANSGENIC RESEARCH
  • 1997-10. Enhanced resistance to two stem borers in an aromatic rice containing a synthetic cryIA(b) gene in MOLECULAR BREEDING
  • 2013-10-27. Combination of site-specific recombination and a conditional selective marker gene allows for the production of marker-free tobacco plants in PLANT CELL, TISSUE AND ORGAN CULTURE (PCTOC)
  • 2007-03-14. Plant sigma factors and their role in plastid transcription in PLANT CELL REPORTS
  • 2004-08. T7 RNA Polymerase-Directed Expression of an Antibody Fragment Transgene in Plastids Causes a Semi-Lethal Pale-Green Seedling Phenotype in TRANSGENIC RESEARCH
  • 2007-11-01. Statistical tools for transgene copy number estimation based on real-time PCR in BMC BIOINFORMATICS
  • 2013-10-20. Chloroplast expression of an HIV envelop-derived multiepitope protein: towards a multivalent plant-based vaccine in PLANT CELL, TISSUE AND ORGAN CULTURE (PCTOC)
  • 2010-09-07. Expression of Bioactive Thymosin Alpha 1 (Tα1) in Marker-free Transgenic Lettuce (Lactuca sativa) in PLANT MOLECULAR BIOLOGY REPORTER
  • 2009-06. High-quality plant DNA extraction for PCR: an easy approach in JOURNAL OF APPLIED GENETICS
  • 2011-09-21. Biosafety of Molecular Farming in Genetically Modified Plants in MOLECULAR FARMING IN PLANTS: RECENT ADVANCES AND FUTURE PROSPECTS
  • 2008-04-03. A protocol for expression of foreign genes in chloroplasts in NATURE PROTOCOLS
  • 2007-05-15. The genetic transformation of plastids in CELL AND MOLECULAR BIOLOGY OF PLASTIDS
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    URI

    http://scigraph.springernature.com/pub.10.1007/s13562-014-0294-2

    DOI

    http://dx.doi.org/10.1007/s13562-014-0294-2

    DIMENSIONS

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