Heterologous expression system in yeast auxotrófica for leucine containing expression vector with check mark defectiva leu2 - D,The process of obtaining this system and use of repeated sequences for multiple integration

摘要

The present invention relates to the construction of expression vector containing the mark defectiva leu2 - D,That allows the selection of cells containing a greater number of copies transform vector integrated into the genome.The scope of the invention comprises an expression system containing a set of vectors and yeast auxotrófica lineage, and the use of repeated sequences of chromosome II.Chromosome III and 5S rDNA of yeast to promote homologous recombination at these loci in the genome of yeast. The presence of these repeated sequences in the vector leads to multiple integration.The line auxotrófica for the amino acid leucine is obtained after genetic modifications in yeast cells to disrupt the gene leu2.The modified cells are used as hosts for the expression of genes present in the vectors containing the check mark leu2 - D.The transformation of the line auxotrófica leu2 with the vectors of the invention enables the recovery of the prototrofia when a high number of copies of the vector is integrated, expressing, that way,The protein encoded by the gene of interest inserted in the vector.