Heterologous expression of biologically active porcine lactoferrin in Pichia Pastoris yeast

The production, expression and biological activities of the recombinant porcine lactoferrin (rpLF) expressed in P. pastoris are discussed. pLF complementary DNA was obtained from total RNA extracted from the mammary gland of a sow in late lactation, amplified using reverse transcription-polymerase chain reaction (RT-PCR) and cloned into the P. pastoris pPIC9 expression vector. The 2100-bp product was analysed by SDS-PAGE and western blotting, resulting in a 80-kDa band of the protein. The recombinant protein at 0.1 mg/ml inhibited the in vitro growth of Escherichia coli isolated from piglets and at 0.05 mg/ml promoted the growth of Lactobacillus casei subsp. casei. Sequence data for the rpLF has been deposited with the EMBL/GenBank Data Libraries under Accession Number AY306198.