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.
Ma
De-ying
DY
Institute of Animal Nutrition, Northeast Agricultural University, Mucai Street 59#, Xiangfang District, Harbin 150030, PR China. mdy296@sohu.com
Liu
Sheng-wang
SW
Han
Zong-xi
ZX
Li
Yi-jin
YJ
Shan
An-shan
AS
D000818:Animals; D002645:Chickens; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000187:drug effects; D008826:Microbial Sensitivity Tests; D017510:Protein Folding; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000494:pharmacology; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000737:chemistry; D017011:Streptococcus suis / Q000187:drug effects; D014018:Tissue Distribution; D023083:beta-Defensins / Q000096:biosynthesis* / Q000378:metabolism
D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D023083:beta-Defensins; C528227:gallinacin-8 protein, chicken; C528228:gallinacin-9 protein, chicken