IBR   13079
INSTITUTO DE BIOLOGIA MOLECULAR Y CELULAR DE ROSARIO
Unidad Ejecutora - UE
congresos y reuniones científicas
Título:
Production of a biologically active recombinant growth hormone from pejerrey
Autor/es:
SCIARA, A.; SOMOZA, G.; ARRANZ, SILVIA EDA
Lugar:
Rosario
Reunión:
Congreso; XLII Reunión anual SAIB; 2006
Resumen:
Since economical equation in fisheries is conditioned on fish growing rates, growthhormone based studies became a key factor in aquaculture development. Currentpejerrey cultures do not provide adequate growing rates for commercial applications.Oral administration of active growth hormone is a valid tool to promote growth inteleost fish. The aim of this work was to obtain an active recombinant pejerrey growthhormone (pjGHr) by the optimization of the solubilization and refolding of pjGHrproduced in a highly efficient prokaryotic system. Hormone cDNA inserted in pET-24vector was expressed in E.coli BL21 CodonPlus. Cells were induced with 2 g/l lactoserendering 0.4 g/l pjGHr with a purity of 83%. To achieve an efficient refolding,inclusion bodies were solubilized in 8M Urea, Tris 100mM pH 10.5. The optimalrefolding condition was obtained by dilution of the protein solution in ten volumes ofTris 40mM pH 10,5 with Arginine 50mM used as an intermolecular interactioninhibitor. Finally the dilution was dialyzed 24 hs against Tris 20mM pH 9 followed by12 hs against sodium bicarbonate 0,05%. Juvenil pejerrey injected with 0.5 μg of pjGHrper gram of body weight exhibited significant increase (P<0.05) of hepatic IGF-ImRNA measured by semi-quantitative RT-PCR. Active pjGHr production is the firststep to achieve an improvement of growing rates of this specie