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

Authors

Lan Li, Northwest University, Xi'anFollow
Chen Zhang, Northwest University, Xi'an
Dan Xu, Northwest University, Xi'anFollow
Michael Schläppi, Marquette UniversityFollow
Zi-Qin Xu, Northwest UniversityFollow

Document Type

Article

Language

eng

Format of Original

12 p.

Publication Date

9-2012

Publisher

Elsevier

Source Publication

Gene

Source ISSN

0378-1119

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.

Comments

Accepted version. Gene, Vol. 506, No. 1 (September 10, 2012): 50-61. DOI. © 2012 Elsevier. Used with permission.

Recommended Citation

Li, Lan; Zhang, Chen; Xu, Dan; Schläppi, Michael; and Xu, Zi-Qin, "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" (2012). Biological Sciences Faculty Research and Publications. 123.
https://epublications.marquette.edu/bio_fac/123