Expression and characterization of recombinant gallinacin-9 and gallinacin-8 in Escherichia coli

Gallinacins (Gal) are antimicrobial peptides that play significant roles in innate immunity in chickens. Two Gal genes—Gal-8 and Gal-9—were cloned and sequenced from chicken liver and tongue, respectively, by reverse transcriptase-polymerase chain reaction (RT-PCR). In addition, the mRNA expression of these genes has been demonstrated across a panel of chicken tissues. It was demonstrated that Gal-9 mRNA was highly expressed in the tongue and small intestine and moderately expressed in the chicken proventriculus, lung, liver, heart, spleen, and thymus. However, Gal-8 mRNA was highly expressed in the chick small intestine and liver, and moderately expressed in the chick tongue, and lung. The recombinant fusion proteins containing Gal-9 or Gal-9 and Gal-8, namely rGal-9 and rGal-9-Gal-8, were produced and purified, respectively. Both rGal-9 and rGal-9-Gal-8 were expressed as insoluble bodies and exhibited the expected antimicrobial activity against Escherichia coli and pathogenic Streptococci suis CAB strain, as determined by the measurement of the inhibition zone and a liquid growth inhibition assay.