Abstract
As a usual response, plants induce/activate various proteins which are thought to be involved in defense mechanisms against the biotic and abiotic stresses they may be confronted with. The novel DUF538 domain containing proteins with unknown functions have been found to be induced/activated in response to different environmental stress stimuli in plants. In order to perform biochemical studies with these new plant stress-responsive proteins, a cDNA containing DUF538 domain was amplified from Celosia cristata full-length leaf expression library using a specific primer set. The isolated cDNA was subsequently expressed in Escherichia coli as a part of maltose-binding fusion protein (MBP-DUF538 construct) and purified at the yield of about 32 mg per liter of cell culture by affinity chromatography without affecting the recombinant bacterial cell growth. The purified fusion product was exogenously applied (10 μg per 4 cm2) on the leaves of Nicotiana tobaccum L. The results revealed that fused DUF538 protein does not induce morphological reposes, but elevates redox enzyme activities including catalase, peroxidase, polyphenol oxidase and phenyalanine ammonia lyase. This is the first time ever time report with respect to the heterologous expression of a plant stress-responsive DUF538 domain that may provide a basis to study its physiological roles and biochemical activities in vitro and in vivo.
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Abbreviations
- MBP:
-
Maltose-binding protein
- DUF:
-
Protein of unknown function
- pfu:
-
Plaque formation unit
- IPTG:
-
Isopropyl-1-thio-ß-D-galactoside
- EDTA:
-
Ethylene diamine tetra-acetic acid
- OD:
-
Optical density
- PVP:
-
Polyvinyl polypyrrolidone
- PAL:
-
Phenylalanine ammonia lyase
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Acknowledgments
The author of this paper is thankful to Research Institute for Fundamental Sciences (RIFS), University of Tabriz, Iran for supporting this work. The author also thanks Department of Biochemistry at PUSA Institue, New Delhi, India for providing the Celosia seeds.
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Gholizadeh, A. Heterologous Expression of Stress-Responsive DUF538 Domain Containing Protein and its Morpho-Biochemical Consequences. Protein J 30, 351–358 (2011). https://doi.org/10.1007/s10930-011-9338-9
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DOI: https://doi.org/10.1007/s10930-011-9338-9