OPTIMIZATION OF RECOMBINANT MAIZE CDKAPRODUCTION IN ESCHERICHIA COLI: ASTATISTICAL APPROACH

AAE MÉNDEZ; SM GARZA-AGUILAR; JM VÁZQUEZ-RAMOS; LB PENA; SM GALLEGO

Cordoba

Congreso; LII Reunión Anual Sociedad Argentina de Investigación en Bioquímica y Biología Molecular; 2016

SAIB

The complex formed by the cyclin dependent kinase A (CDKA)and cyclin D is responsible for the G1-Stransition of cell cycle. Oxidative post-translational modifications of these proteins block cell cycle progression, and thus contributing in plant growth inhibition during abiotic stress. As part of a project to determine possible sites of CDKA protein oxidation, CDKA from ZeamaysL (maize) was cloned and expressed in E. coli. The present study describes the optimization of the production of the protein using as statistical approach: Response Surface Methodology. A 24 full factorial central composite rotary design for four independent variables, each at five levels was employed to fit asecond order polynomial model which indicated that 30 experiments were required for this procedure. Level 0 of variables used was 26°C, 0,6 OD 600nm before induction, 0,75 mM isopropyl β-D-1-thiogalactopyranoside(IPTG) concentration and 5 h post-induction time. An optimization in CDKA production in the soluble fraction was achieved. Results obtained are not valid for all recombinant proteins.