Hostname: page-component-8448b6f56d-mp689 Total loading time: 0 Render date: 2024-04-16T11:34:17.938Z Has data issue: false hasContentIssue false
  • English
  • Français

Cloning and expression of Ma duck interleukin-18 mature protein gene and biological activity detection

Published online by Cambridge University Press:  13 February 2008

Chen Hong-Ying ,
Cui Bao-An ,
Xia Ping-An ,
Li Xin-Sheng ,
Yang Ming-Fan  and
Zheng Lan-Lan
Chen Hong-Ying
Affiliation:
College of Animal Husbandry and Veterinary Medicine, Henan Agricultural University, Zhengzhou 450002, China Animal Food Safety Key Laboratory, Henan Province, Zhengzhou 450002, China
Cui Bao-An*
Affiliation:
College of Animal Husbandry and Veterinary Medicine, Henan Agricultural University, Zhengzhou 450002, China Animal Food Safety Key Laboratory, Henan Province, Zhengzhou 450002, China
Xia Ping-An
Affiliation:
College of Animal Husbandry and Veterinary Medicine, Henan Agricultural University, Zhengzhou 450002, China
Li Xin-Sheng
Affiliation:
College of Animal Husbandry and Veterinary Medicine, Henan Agricultural University, Zhengzhou 450002, China
Yang Ming-Fan
Affiliation:
College of Animal Husbandry and Veterinary Medicine, Henan Agricultural University, Zhengzhou 450002, China Animal Food Safety Key Laboratory, Henan Province, Zhengzhou 450002, China
Zheng Lan-Lan
Affiliation:
College of Animal Husbandry and Veterinary Medicine, Henan Agricultural University, Zhengzhou 450002, China Animal Food Safety Key Laboratory, Henan Province, Zhengzhou 450002, China
*
*Corresponding author. E-mail: baoancui@henau.edu.cn
Get access Rights & Permissions [Opens in a new window]

Abstract

Duck interleukin (IL)-18 mature protein gene was amplified by reverse transcription-polymerase chain reaction (RT-PCR) from total RNA extracted from Ma duck (Tadorna ferruginea) splenocytes. The PCR product was cloned into pGEM-T Easy vector for sequencing. The result revealed that the nucleotide sequence of duck IL-18 mature protein gene (mDuIL-18) consisted of a 513 bp band. A prokaryotic plasmid of mDuIL-18, pQE30-mDuIL18, was obtained by subcloning the encoding region of the DuIL-18 mature peptide into pQE30. pQE30-mDuIL18 transformed Escherichia coli M15. The expression of mDuIL-18 gene was identified by SDS-PAGE and Western blotting. Its molecular weight was 19.76 kDa, and could be specifically recognized by rabbit sera to chicken IL-18. The expressed products existed as inclusion bodies. After being degenerated, then renatured, the activities of the inclusion bodies were detected by methyl thiazolyl tetrazolium (MTT) assay. In ducks injected intramuscularly with mDuIL-18 protein (150 ng or 200 ng per duck) and Avian influenza virus (AIV) vaccine 2 weeks after immunization, the average titres of haemagglutination inhibition (HI) antibodies to AIV reached 7.5–7.7 log2, while those of HI antibody ranged between 6.3 and 6.6 log2 in ducks vaccinated with AIV vaccine only or with 100 ng mDuIL-18 and AIV vaccine. The results clearly showed that 150 ng mDuIL-18 per duck strengthened the in vivo immune responses induced by the inactivated oil emulsion AIV vaccine.

Keywords

biological activitycloningduck interleukin-18expressionmature protein gene
Type
Research Article
Information
Chinese Journal of Agricultural Biotechnology , Volume 4 , Issue 3 , December 2007 , pp. 207 - 212
Copyright
Copyright © China Agricultural University and Cambridge University Press 2007

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

Footnotes

First published in Journal of Agricultural Biotechnology 2007, 15(4): 584–588

References

Bokarewa, M and Hultgren, O (2005) Is interleukin-18 useful for monitoring rheumatoid arthritis? Scandinavian Journal of Rheumatology 34(6): 433–436.CrossRefGoogle ScholarPubMed
Chen, HY, Cui, BA, Huang, QY, Li, XS and Wang, XB (2006) Cloning and sequencing of the chicken interleukin-18. Chinese Journal of Preventive Veterinary Medicine 28(1): 48–50 (in Chinese with English abstract).Google Scholar
Degen, WG, van Zuilekom, HI, Scholtes, NC, van Daal, N and Schijns, VE (2005) Potentiation of humoral immune responses to vaccine antigens by recombinant chicken IL-18 (rChIL-18). Vaccine 23(33): 4212–4218.CrossRefGoogle ScholarPubMed
Dinarello, CA and Fantuzzi, G (2003) Interleukin-18 and host defense against infection. Journal of Infectious Diseases 187, S370–S384.Google Scholar
Gobel, TW, Schneider, K, Schaerer, B, et al. (2003) IL-18 stimulates the proliferation and IFN-gamma release of CD4+ T cells in the chicken: Conservation of a Th1-like system in a nonmammalian species. Journal of Immunology 171(4): 1809–1815.CrossRefGoogle Scholar
Gracie, JA, Robertson, SE and McInnes, IB (2003) Interleukin-18. Journal of Leukocyte Biology 73, 213–224.Google Scholar
Gu, Y, Kuida, K, Tsutsui, H, et al. (1997) Activation of interferon-γ inducing factor mediated by interleukin-1β converting enzyme. Science 275: 5297206–5297209.CrossRefGoogle ScholarPubMed
Hu, JD, Zhao, HK and Cui, ZZ (2006) Construction of eukaryotic expressing plasmid of chicken interleukin 18 mature protein gene and its expression in chick embryonic fibroblasts (CEF). Acta Veterinaria et Zootechnica Sinica 37(1): 91–94 (in Chinese with English abstract).Google Scholar
Jia, LJ, Liu, XF, Zhang, YM, Peng, DX and Zhang, RK (2004) Effect of rPPV-IFN as immunopotentiator on active immunization induced by H5 subtype avian influenza vaccines in chickens. Journal of Agricultural Biotechnology 12(4): 427–430 (in Chinese with English abstract).Google Scholar
Puehler, F, Gobel, T, Breyer, U, Ohnemus, A, Staeheli, P and Kaspers, B (2003) A sensitive bioassay for chicken interleukin-18 based on the inducible release of preformed interferon-gamma. Journal of Immunological Methods 274(1–2): 229–232.CrossRefGoogle ScholarPubMed
Sambrook, J, Fritsch, EF and Maniatis, T (1989) Molecular Cloning: A Laboratory Manual. 2nd ed. New York: Cold Spring Harbor Laboratory Press.Google Scholar
Schneider, K and Puehler, F (2000) cDNA Cloning of biologically active chicken interleukin-18. Journal of Interferon and Cytokine Research 20: 879–883.Google Scholar
Shinoda, M, Wakabayashi, G, Shimazu, M, et al. (2006) Increased serum and hepatic tissue levels of interleukin-18 in patients with fulminant hepatic failure. Journal Gastroenterology and Hepatology 21(11): 1731–1736.CrossRefGoogle ScholarPubMed
Thong-Ngam, D, Tangkijvanich, P, Lerknimitr, R, Mahachai, V, Theamboonlers, A and Poovorawan, Y (2006) Diagnostic role of serum interleukin-18 in gastric cancer patients. World Journal of Gastroenterology 12(28): 4473–4477.CrossRefGoogle ScholarPubMed
Yano, A, Nakao, K, Sarai, A, et al. (2005) Elevated serum interleukin-18 levels might reflect the high risk of hospitalization in patients on peritoneal dialysis. Nephrology (Carlton) 10(6): 576–582.CrossRefGoogle ScholarPubMed