Bioimpacts. 2017;7(4):247-254.
doi: 10.15171/bi.2017.29
PMID: 29435432
PMCID: PMC5801536
Scopus id: 85040972974
WOS: 000423937100006
  Abstract View: 405
  PDF Download: 339
  Full Text View: 126

Original Research

Efficient and stable transformation of Dunaliella pseudosalina by 3 strains of Agrobacterium tumefaciens

Jaber Dehghani 1,2, Ali Movafeghi 1 * , Abolfazl Barzegari 2,3, Jaleh Barar 2 * ORCiD

1 Department of Plant Biology, Faculty of Natural Science, University of Tabriz, 29th Bahman Blvd, Tabriz, Iran
2 Research Center for Pharmaceutical Nanotechnology, Tabriz University of Medical Science, Tabriz, Iran
3 School of Advanced Biomedical Sciences, Tabriz University of Medical Science, Daneshgah street, Tabriz, Iran

Abstract

Introduction: Several platforms including mammalian, plant and insect cells as well as bacteria, yeasts, and microalgae are available for the production of recombinant proteins. Low efficiency of delivery systems, extracellular and intracellular degradation of foreign genes during transformation, difficulties in targeting and importing into the nucleus, and finally problems in integration into nuclear genome are the most bottlenecks of classical plasmids for producing recombinant proteins. Owing to high growth rate, no common pathogen with humans, being utilized as humans’ food, and capability to perform N-glycosylation, microalgae are proposed as an ideal system for such biotechnological approaches. Here, Agrobacterium tumefaciens is introduced as an alternative tool for transformation of the microalga Dunaliella pseudosalina.
Methods: The transformation of gfp gene into the D. pseudosalina was evaluated by three strains including EHA101, GV3301 and GV3850 of A. tumefaciens. The integrating and expression of gfp gene were determined by PCR, RT-PCR, Q-PCR and SDS-PAGE analyses.
Results: The T-DNA of pCAMBIA1304 plasmid was successfully integrated into the genome of the microalgal cells. Although all of the strains were able to transform the algal cells, GV3301 possessed higher potential to transform the microalgal cells in comparison to EHA101 and GV3850 strains. Moreover, the stability of gfp gene was successfully established during a course of two months period in the microalgal genome.
Conclusion: Agrobacterium is introduced as a competent system for stable transformation of Dunaliella strains in order to produce eukaryotic recombinant proteins.
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Submitted: 29 May 2017
Revised: 30 Aug 2017
Accepted: 05 Sep 2017
First published online: 18 Sep 2017
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