title: Heterologous expression of stress-responsive DUF538 domain containing protein and its morpho-biochemical consequences.
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 cm(2)) 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.
authors.affiliation: Research Institute for Fundamental Sciences (RIFS), University of Tabriz, Iran. aghz_bioch@yahoo.co.in
authors.forename: Ashraf
authors.initials: A
authors.lastname: Gholizadeh
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D034762:Celosia / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli; D008969:Molecular Sequence Data; D010084:Oxidation-Reduction; D018384:Oxidative Stress / Q000187:drug effects*; D010088:Oxidoreductases / Q000378:metabolism; D010650:Phenylalanine Ammonia-Lyase / Q000378:metabolism; D010766:Phosphorylation; D018515:Plant Leaves / Q000187:drug effects / Q000378:metabolism; D010940:Plant Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology*; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology*; D020539:Sequence Analysis, Protein; D014026:Nicotiana / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism
chemical_list: D010940:Plant Proteins; D011994:Recombinant Proteins; D010088:Oxidoreductases; D010650:Phenylalanine Ammonia-Lyase