SciELO - Scientific Electronic Library Online

 
vol.27 issue2Preliminary evaluation and updating of the productive losses to improve the profitability of FinlayA proposal for input classification for inventory management in the biopharmaceutical industry. Study case in the Molecular Immunology Center author indexsubject indexarticles search
Home Pagealphabetic serial listing  

Services on Demand

Journal

Article

Indicators

  • Have no cited articlesCited by SciELO

Related links

  • Have no similar articlesSimilars in SciELO

Share


Vaccimonitor

Print version ISSN 1025-028XOn-line version ISSN 1025-0298

Abstract

GONZALEZ-CRESPO, Annia; HARDY-SOSA, Anette  and  SOSA-ESPINOSA, Angela E.. Rapid verification method of Escherichia coli mutants used in genetic engineering. Vaccimonitor [online]. 2018, vol.27, n.2, pp.45-50. ISSN 1025-028X.

For recombinant DNA methodologies, molecular biologists use Escherichia coli mutants acquired from commercial kits or specialized microbial strain collections. These mutants are routinely conserved by successive passages or in poorly-characterized strain banks. A failure to follow the seed lot system (reference seed lot and working seed lot) and the lack of systematic controls, can lead to the loss of original strain characteristics and affect the quality of experimental results. On the other hand, the dynamic of research laboratories makes impractical to carry out extensive verifications related to the work with microbial collections. The aim of this article is to propose a single-assay method for genetic purity and stability evaluation of multiple E. coli mutants simultaneously. For the design of specific culture media, selection criteria were defined. Serial dilutions of cultures in Luria Bertani medium were made, and track dilution was used as seeding method. The results show how a mutated metabolic pathway-oriented design permit the verification of several strains in a single trial. A criterion about culture purity and genetic stability could be obtained after combining specific media. This alternative allows for a rapid evaluation of strain key genetic markers for recombinant DNA methodologies

Keywords : Escherichia coli; genotype; recombinant DNA.

        · abstract in Spanish     · text in Spanish     · Spanish ( pdf )

 

Creative Commons License All the contents of this journal, except where otherwise noted, is licensed under a Creative Commons Attribution License