Successful production of the potato antimicrobial peptide Snakin‐1 in baculovirus‐infected insect cells and development of specific antibodies.

ALMASIA, NATALIA INÉS; MOLINARI, MARÍA PAULA; MARONICHE, GUILLERMO ANDRÉS; NAHIRÑAK, VANESA; BARRIOS BARÓN, MARÍA PILAR; TABOGA, OSCAR ALBERTO; VAZQUEZ ROVERE, CECILIA

Bordeaux

Congreso; 4èmes Journées GDR MuFoPAM; 2017

Snakin-1 is a broad-spectrum antimicrobial cysteine-rich peptide isolated from Solanumtuberosum. Its biotechnological potential has been already recognized since it exhibits invivo antifungal and antibacterial activity. Most attempts to produce StSN1, or homologouspeptides, in a soluble native state using bacterial, yeast or synthetic expression systems havepresented production bottlenecks such as insolubility, misfolding or low yields. In this work,we successfully expressed a recombinant StSN1 in Spodoptera frugiperda insect cells byoptimizing several of the parameters for its expression in the baculovirus expression system.The recombinant peptide was soluble and was present in the nuclear fraction of infected Sf9cells. An optimized purification procedure allowed the production of rSN1 that was used forimmunization of mice, which gave rise to polyclonal antibodies that detect the native proteinin tissue extracts of both agroinfiltrated plants and stable transgenic lines. We suggest theuse of the baculovirus system for high-level expression of Snakin/GASA peptides, forbiological assays, structural and functional analysis and antibody production, as an importantstep to both elucidate their accurate physiological role and to deepen the study of theirbiotechnological uses.