Cluster Transfronterizo Biotecnológico

CICA-INIBIC. Procedure for detection of recombinant homologous recombination in vivo by analysing recombinant gfp forms

Project Description

The Yeast Gene Expression (Levagen) and Endocrine, Nutritional and Medical Physiopathology (FENM) groups are looking for companies interested in licensing the system for the identification, with fluorescence microscopy, of genome rearrangements (recombination) caused by different conditions. They use yeasts that express fluorescent proteins from different cellular locations.

Description of the Area of Activity / Technology

This technology in the current state of development, allows to identify changes originated by mutations or external chemical agents and/or drugs, which cause gene reorganization, using yeast as a model organism and without the need for any type of cellular processing. Only visualization by fluorescence microscopy. A great advantage over studies that apply PCR techniques, is here the cells are visualized directly after culture, while the PCR analysis requires the processing of the samples, the PCR itself and electrophoresis.

On the other hand, with our technique we can identify recombination in individual cells that could go unnoticed, if their frequency is low, in a PCR with the DNA of several cells. In the case of the study of recombination by classical genetic techniques, it is necessary to detect cell growth in selective plates, which involves a waste of time (usually 2-3 days) and culture materials.

The technology is applicable and patented for use in any type of eukaryotic cell. Therefore in these cases it is not necessary to make DNA extractions or PCRs and also allows differentiating the direction of recombination as the cytosolic or nucleolar bloom is lost. It includes two yeast strains to have a positive and a negative control in the studies to which it is applied. The technology allows the development of a specific image analysis software.