ISSN: 0973-7510

E-ISSN: 2581-690X

Cesar Alvarez-Mejia1, Gustavo Hernandez-Guzman2, Everardo Curiel-Quesada3 and Rodolfo Marsch4
1Department of Plant Genetic Engineering, Cinvestav-IPN, U.-Irapuato, K.M. 9 Road Irapuato-Leon Irapuato, Gto., 36500, Mexico.
2Department of Life Sciences, University of Guanajuato, El Copal K.M. 9 Road Irapuato-Silao; Irapuato, Gto., 36500, Mexico.
3Department of Biochemistry, National Polytechnic Institute, National School of Biological Sciences, Carpio y Plan de Ayala S/N, Colonia Santo Tomás, 11340 México, D.F.,
4Department of Biotechnology and Bioengineering, Cinvestav. Avenue I.P.N. 2508, Colonia San Pedro Zacatenco, 07360. Mexico, D.F. Mexico.
J Pure Appl Microbiol. 2013;7(3):1743-1748
© The Author(s). 2013
Received: 08/01/2013 | Accepted: 27/02/2013 | Published: 30/09/2013
Abstract

The reporter gene cat can be used to determine promoter activation and to seek genes related to pathogenicity, where the promoter becomes tagged. The estimation of promoter expression in these cases can be evaluated by three approaches: i) determination of relative quantity of the mRNA; ii) quantification of protein (chloramphenicol acetyl-transferase); or iii) determination of specific enzymatic activity. However the ultimate result for cat expression in a cell is to endow resistance against chloramphenicol. In this work we use the cell density measure with chloramphenicol antibiotic as an analysis of the promoter regulation and strength during a study of Pseudomonas syringae pv. maculicola mutant screening. We found that the promoter expression level modifies accordingly the cell density in liquid media and also the colony size in solid media at defined times. We propose the determination of cell density in liquid media supplemented with a constant concentration of chloramphenicol to estimate not only the expression conditions of a promoter tagged with cat, but also the expression intensity in those conditions.

Keywords

Promoter expression, cat gene, Pseudomonas syringae pv. maculicola M2, Tn5-cat1

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