Clonagem e expressão do gene da glicoproteína "E" do vírus da doença de Aujeszky em sistema de expressão Pichia pastoris

Abstract

Aujeszky's disease (AD) is caused by Suid Herpesvirus 1 (SHV1) and affects a wide variety of animals, the pig being the most important host. The DA has caused considerable direct and indirect economic losses to the pig industry worldwide, from the impact on production. The use of control measures using diagnostic test able to differentiate infected animals from vaccinated animals and removal of positive animals in the squad is key to controlling the spread of the virus. The use of vaccines for the gE gene deleted of SHV1 has been used to decrease clinical signs and virus transmission. Although some epitopes gE gene has already been cloned into the Pichia pastoris expression system, relative to the portion extramembranares epitopes using synthetic sequence (gEs) yeast codon optimized for this has not yet been cloned in this system. In this work this sequence was cloned and expressed in P. pastoris, which produced the recombinant antigen gEr. The gEs was cloned in strains X-33 and KM71H this yeast. The two strains were induced to express cloned gEr. The recombinant protein was detected in both clones by colony blot and visualized on SDS-PAGE gel, revealing a protein of approximately 50 kDa.