Heterologous protein expression in the methylotropic yeast Pichia methanolica.

LUCERO AA, MANZUR MJ, VILLARREAL R, CIUFFO GM

Huerta Grande, Cordoba, Argentina

Congreso; Reunión anual de la Sociedad de Biología de Cuyo; 2007

Recombinant proteins have become of wide use in laboratories as well as clinical applications. Different expression systems have been used for these purpose, becoming a common and frequent tool in Molecular Laboratories.  In the past years, the methylotropic yeast Pichia methanolica has developed into a highly successful system for the production of a wide variety of heterologous proteins.  This eukaryotic organism has several characteristics that suggest it is well-suited for expression of recombinant proteins. The conditions for protein expression system in the methylotropic yeast Pichia methanolica, were set up for a fragment of Angiotensin II AT2 receptor.   Starting from the fragment (585 bp) subcloned in pGEMT easy vector; the C-terminal fragment was subcloned at the three open reading frames of pMET a vector. We achieved the expression, secretion and identification of the AT2 fragment into the growth media of our recombinant protein. Yeast combines the easy of genetic manipulation and fermentation of a microbial organism with the capability of secrete and to modify proteins according to a general eukaryotic scheme.