title: Expression of recombinant EARLI1, a hybrid proline-rich protein of Arabidopsis, in Escherichia coli and its inhibition effect to the growth of fungal pathogens and Saccharomyces cerevisiae.
abstract: EARLI1 is an Arabidopsis gene with pleiotropic effects previously shown to have auxiliary functions in protecting plants against freezing-induced cellular damage and promoting germinability under low-temperature and salinity stresses. Here we determined whether recombinant EARLI1 protein has anti-fungal activity. Recombinant EARLI1 protein lacking its signal peptide was produced in Escherichia coli BL21(DE3) using isopropyl β-d-1-thiogalactopyranoside (IPTG) induction and the prokaryotic expression vector pET28a. Expression of EARLI1 was analyzed by Western blotting and the protein was purified using affinity chromatography. Recombinant EARLI1 protein was applied to fungal cultures of Saccharomyces cerevisiae, Botrytis cinerea and Fusarium oxysporum, and membrane permeability was determined using SYTOX green. Full-length EARLI1 was expressed in S. cerevisiae from the GAL1 promoter using 2% galactose and yeast cell viability was compared to control cells. Our results indicated that application of recombinant EARLI1 protein to B. cinerea and F. oxysporum could inhibit the growth of the necrotrophic fungi. Besides, addition of the recombinant protein to liquid cultures of S. cerevisiae significantly suppressed yeast growth and cell viability by increasing membrane permeability, and in vivo expression of the secreted form of EARLI1 in S. cerevisiae also had a remarkable inhibition effect on the growth of yeast cells.
authors: Key Laboratory of Resource Biology and Biotechnology in Western China (Ministry of Education), Provincial Key Laboratory of Biotechnology, Institute of Life Sciences, Northwest University, Xi'an, Shaanxi 710069, People's Republic of China.
authors: Lan
authors: L
authors: Li
authors: Chen
authors: C
authors: Zhang
authors: Dan
authors: D
authors: Xu
authors: Michael
authors: M
authors: Schläppi
authors: Zi-Qin
authors: ZQ
authors: Xu
mesh_terms: D000595:Amino Acid Sequence; D000935:Antifungal Agents / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology*; D052179:Antigens, Plant / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology; D017360:Arabidopsis / Q000235:genetics / Q000502:physiology; D029681:Arabidopsis Proteins / Q000737:chemistry / Q000235:genetics* / Q000494:pharmacology*; D001483:Base Sequence; D020171:Botrytis / Q000187:drug effects / Q000254:growth & development / Q000472:pathogenicity; D002352:Carrier Proteins / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology; D002463:Cell Membrane Permeability / Q000187:drug effects; D018744:DNA, Plant / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005670:Fusarium / Q000187:drug effects / Q000254:growth & development / Q000472:pathogenicity; D015870:Gene Expression; D017343:Genes, Plant; D005822:Genetic Vectors; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000254:growth & development; D017386:Sequence Homology, Amino Acid
chemical_list: D000935:Antifungal Agents; D052179:Antigens, Plant; D029681:Arabidopsis Proteins; D002352:Carrier Proteins; D018744:DNA, Plant; C419264:EARLI1 protein, Arabidopsis; D010940:Plant Proteins; D011994:Recombinant Proteins; C511920:lipid transfer proteins, plant