title: Prokaryotic expression of chicken interferon-γ fusion protein and its effect on expression of poultry heat shock protein 70 under heat stress.
abstract: Interferons have attracted considerable attention due to their vital roles in the host immune response and low induction of antibiotic resistance. In this study, total RNA was extracted from spleen cells of chicken embryos inoculated with Newcastle disease vaccine, and the full-length chicken interferon-γ (ChIFN-γ) gene was amplified by RT-PCR. The full complementary DNA sequence of the ChIFN-γ gene was 495 bp long and was cloned into the prokaryotic expression vector pProEX™HTb. The plasmid was transformed into Escherichia coli DH5α and the expression of ChIFN-γ was induced by isopropyl β-D-1-thiogalactopyranoside. Sodium dodecyl sulfate - polyacrylamide gel electrophoresis and Western blot results showed the expressed fusion protein had a molecular weight of approximately 18 kDa and was recognized by an anti-His mAb. Moreover, ChIFN-γ was found to demonstrate anti-viral activity in vitro. To test the in vivo function of ChIFN-γ in broilers under heat stress, a total of 100 broilers were randomly assigned to either a control group or a treated group, in which they were hypodermically injected with recombinant ChIFN-γ. Results demonstrated ChIFN-γ affects the messenger RNA expression levels of heat shock protein 70 (HSP70) in the heart and lung tissues, and decreases the concentration of HSP70 in serum. Therefore, we conclude recombinant ChIFN-γ can reduce heat stress to some extent in vivo.
authors.affiliation: College of Animal Science and Technology, Northeast Agricultural University, Harbin, China.
authors.forename: Jinhua
authors.initials: J
authors.lastname: Sun
authors.affiliation: College of Animal Science and Technology, Northeast Agricultural University, Harbin, China.
authors.forename: Yinglin
authors.initials: Y
authors.lastname: Chen
authors.affiliation: College of Animal Science and Technology, Northeast Agricultural University, Harbin, China.
authors.forename: Feiyue
authors.initials: F
authors.lastname: Qin
authors.affiliation: College of Animal Science and Technology, Northeast Agricultural University, Harbin, China.
authors.forename: Xueting
authors.initials: X
authors.lastname: Guan
authors.affiliation: College of Animal Science and Technology, Northeast Agricultural University, Harbin, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Xu
authors.affiliation: College of Animal Science and Technology, Northeast Agricultural University, Harbin, China.
authors.forename: Liangmei
authors.initials: L
authors.lastname: Xu
mesh_terms: D000818:Animals; D000998:Antiviral Agents; D002642:Chick Embryo; D002645:Chickens; D015870:Gene Expression; D018840:HSP70 Heat-Shock Proteins / Q000097:blood / Q000235:genetics* / Q000378:metabolism*; D018882:Heat Stress Disorders / Q000517:prevention & control; D018869:Heat-Shock Response / Q000235:genetics* / Q000502:physiology*; D007371:Interferon-gamma / Q000008:administration & dosage / Q000235:genetics* / Q000378:metabolism* / Q000494:pharmacology; D008168:Lung / Q000378:metabolism; D009206:Myocardium / Q000378:metabolism; D012313:RNA / Q000235:genetics* / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000008:administration & dosage / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D013154:Spleen / Q000166:cytology / Q000196:embryology / Q000378:metabolism
chemical_list: D000998:Antiviral Agents; D018840:HSP70 Heat-Shock Proteins; D011993:Recombinant Fusion Proteins; D012313:RNA; D007371:Interferon-gamma
keywords: biological activity; chicken interferon-γ; heat shock protein 70; heat stress; prokaryotic expression