Construction of an expression vector pVLT-EGFP can be expressed in Azospirillum brasilense

doi:10.11924/j.issn.1000-6850.2014-2386

Abstract

Abstract: In order to study the colonization of Azospirillum brasilense in plants, an Escherichia coli-A. brasilense shuttle vector pVLT-EGFP can be expressed in A. brasilense was constructed by integrating the enhanced green fluorescent protein (EGFP) gene of pcDNA3.1-EGFP into the broad host range expression plasmid pVLT-33. The expression vector was transformed into A. brasilense by electroporation methods and the expression of EGFP gene was observed under fluorescent microscope. Using IPTG as an inducing agent, the effect of different time and temperature of IPTG on the expression of pVLT-EGFP was analyzed and the results indicated that the optimal condition for expression was at the temperature of 30℃. The highest expression level of EGFP gene was detected 9 h after inducing. This study enriched A. brasilense genetic transformation system and provided a robust tool to study the colonization of A. brasilense and growth-promoting effect on plants.

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