Molecular Cloning and Sequence Analysis of the Gene Encoding Interferon Alpha of the Giant Panda (Ailuropoda melanoleuca)

Abstract:

nterferon-alpha (IFN-a) is a kind of cytokines that share antiviral, as well as immunomodulatory, and antiproliferative effects on cell functions. In this report, the cDNA for Ailuropoda melanoleucas interferon alpha was cloned from the ConA-stimulated giant panda peripheral blood mononuclear cells (PBMCs) by RT-PCR. Bioinformatics analysis was performed to predict the characteristics of this gene. Sequencing revealed that the fragment was composed of 495 nucleotides, intronless, encoding a mature polypeptide with 164 amino acids and exhibiting a molecular mass of 18.15 kDa. The analysis of the functional sites and antigenic determinants demonstrated that this protein has 27 functional sites and 9 antigenic determinants, And possesses typical characteristics of interferon alpha, beta and delta family. Compared with 10 corresponding IFN-α sequences. It revealed that the GpIFN-a gene had a close evolutionary relationship with mammalians IFN-a. Phylogenetic tree based on nucleotide sequences showed that giant panda, ferret, dog and cat clustered together and evolved into a distinct phylogenetic lineage. In conclusion, all the data and consequences will provide a basis for further functional study of the Ailuropoda melanoleuca IFN-a.

• Periodical: Applied Mechanics and Materials (Volumes 195-196)
• Pages: 370-379
• DOI: https://doi.org/10.4028/www.scientific.net/AMM.195-196.370
• Online since: August 2012
• Authors: Yue Yi, Cheng Dong Wang, Zhi Wen Xu, De Sheng Li, Ling Zhu, Feng Qin Li, Feng Gu, Wan Zhu Guo, Cheng Dong Wang, De Sheng Li
• Keywords: Ailuropoda melanoleuca, Clone, Interferon Alpha, Sequence Analysis, Structure Analysis

References

[1] W. Feng and G. Li, The saving of giant panda, Published by Sichuan Science and Technology Publishing Company, pp.113-129, 2000. (in chinese).

[2] D. Liu, Z. Wang, H. Tian, C. Yu, G. Zhang, R. Wei, and H. Zhang, Behavior of giant pandas (Ailuropoda melanoleuca) in captive conditions: Gender differences and enclosure effects, Zoo Biology, vol. 22, pp.77-82, 2003. (in chinese).

DOI: 10.1002/zoo.10076

[3] J. Zhang, P. Daszak, H. Huang, G. Yang, A. Kilpatrick, and S. Zhang, Parasite threat to panda conservation, EcoHealth, vol. 5, pp.6-9, 2008. (in chinese).

DOI: 10.1007/s10393-007-0139-8

[4] W. Stewart and J. Vil ek, The interferon system: Springer-Verlag New York, (1979).

[5] P. Familletti, R. McCandliss, and S. Pestka, Production of high levels of human leukocyte interferon from a continuous human myeloblast cell culture, Antimicrobial agents and chemotherapy, vol. 20, p.5, (1981).

DOI: 10.1128/aac.20.1.5

[6] P. Gray, D. Leung, D. Pennica, E. Yelverton, R. Najarian, C. Simonsen, R. Derynck, P. Sherwood, D. Wallace, and S. Berger, Expression of human immune interferon cDNA in E. coli and monkey cells, (1982).

DOI: 10.1038/295503a0

[7] K. DEVOS, F. DUERINCK, K. VAN AUDENHOVE, and W. FIERS, Cloning and expression of the canine interferon-γ gene, Journal of interferon research, vol. 12, pp.95-102, (1992).

DOI: 10.1089/jir.1992.12.95

[8] K. Vandenbroeck, R. Dijkmans, A. Aerschot, and A. Billiau, Engineering by PCR-based exon amplification of the genomic porcine interferon-gamma DNA for expression in Escherichia coli, Biochemical and biophysical research communications, vol. 180, pp.1408-1415, (1991).

DOI: 10.1016/s0006-291x(05)81353-1

[9] M. DIGBY and J. LOWENTHAL, Cloning and expression of the chicken interferon-γ gene, Journal of Interferon & Cytokine Research, vol. 15, pp.939-945, (1995).

DOI: 10.1089/jir.1995.15.939

[10] X. Tan, Y. Tang, Y. Yang, H. Song, and Y. Zhang, Gene cloning, sequencing, expression and biological activity of giant panda (Ailuropoda melanoleuca) interferon-[alpha], Molecular immunology, vol. 44, pp.3061-3069, (2007).

DOI: 10.1016/j.molimm.2006.12.017

[11] Z. Wang, Animal and human cell genetic techniques: chengdu: Sichuan University Press, 1987. (in chinese).

[12] J. Banchereau, F. Briere, C. Caux, J. Davoust, S. Lebecque, Y. Liu, B. Pulendran, and K. Palucka, Immunobiology of dendritic cells, Annual review of immunology, vol. 18, pp.767-811, (2000).

DOI: 10.1146/annurev.immunol.18.1.767

[13] N. Kadowaki, S. Antonenko, J. Lau, and Y. Liu, Natural interferon α/β–producing cells link innate and adaptive immunity, The Journal of experimental medicine, vol. 192, p.219, (2000).

DOI: 10.1084/jem.192.2.219

[14] B. Charleston, M. Fray, S. Baigent, B. Carr, and W. Morrison, Establishment of persistent infection with non-cytopathic bovine viral diarrhoea virus in cattle is associated with a failure to induce type I interferon, Journal of General Virology, vol. 82, p.1893, (2001).

DOI: 10.1099/0022-1317-82-8-1893

[15] J. Chinsangaram, M. Piccone, and M. Grubman, Ability of foot-and-mouth disease virus to form plaques in cell culture is associated with suppression of alpha/beta interferon, Journal of virology, vol. 73, p.9891, (1999).

DOI: 10.1128/jvi.73.12.9891-9898.1999

[16] E. ALBINA, C. CARRAT, and B. CHARLEY, Short Communication: Interferon-α Response to Swine Arterivirus (PoAV), the Porcine Reproductive and Respiratory Syndrome Virus, Journal of Interferon & Cytokine Research, vol. 18, pp.485-490, (1998).

DOI: 10.1089/jir.1998.18.485

[17] J. Plachy, K. Weining, and M. Kremmer, Proctive effectsof type l and typeⅡinterferon toward Rouse Sarcoma virus-induced tumors in chicken, Virology, vol. 256, pp.85-91, (1999).

DOI: 10.1006/viro.1999.9602

[18] P. Marcus, L. Heide, and M. Sekellick, Interferon action on avian viruses. I. Oral administration of chicken interferon-alpha ameliorates Newcastle disease, Journal of Interferon & Cytokine Research, vol. 19, pp.881-885, (1999).

DOI: 10.1089/107999099313406

[19] U. Schultz and F. Chisari, Recombinant duck interferon gamma inhibits duck hepatitis B virus replication in primary hepatocytes, Journal of virology, vol. 73, p.3162, (1999).

DOI: 10.1128/jvi.73.4.3162-3168.1999

[20] W. Feng, R. Wang, S. Zhong, Z. Ye, X. Cui, and J. Zeng, Analysis on the dead cause of the anatomical carcass of giant panda (Ailuropoda melanoleuca), A study on breeding and diseases of the giant panda, pp.244-248. (in chinese).

[21] H. Tjalsma, A. Bolhuis, J. Jongbloed, S. Bron, and J. van Dijl, Signal peptide-dependent protein transport in Bacillus subtilis: a genome-based survey of the secretome, Microbiology and Molecular Biology Reviews, vol. 64, p.515, (2000).

DOI: 10.1128/mmbr.64.3.515-547.2000

[22] G. von Heijne, The signal peptide, Journal of Membrane Biology, vol. 115, pp.195-201, (1990).

[23] R. Wonderling, T. Powell, S. Baldwin, T. Morales, S. Snyder, K. Keiser, S. Hunter, E. Best, M. McDermott, and M. Milhausen, Cloning, expression, purification, and biological activity of five feline type I interferons, Veterinary immunology and immunopathology, vol. 89, pp.13-27, (2002).

DOI: 10.1016/s0165-2427(02)00188-5

[24] R. Devos, H. Cheroutre, Y. Taya, W. Degrave, H. Van Heuverswyn, and W. Fiers, Molecular cloning of human immune interferon cDNA and its expression in eukaryotic cells, Nucleic acids research, vol. 10, p.2487, (1982).

DOI: 10.1093/nar/10.8.2487

[25] E. Weerapana and B. Imperiali, Asparagine-linked protein glycosylation: from eukaryotic to prokaryotic systems, Glycobiology, vol. 16, p. 91R, (2006).

DOI: 10.1093/glycob/cwj099

[26] R. Huby, R. Dearman, and I. Kimber, Why are some proteins allergens?, Toxicological Sciences, vol. 55, p.235, (2000).

DOI: 10.1093/toxsci/55.2.235

[27] L. Xu, B. Zeng, R. Peng, F. Zou, and B. Yue, Molecular cloning and sequence analysis of the gene encoding interleukin-6 of the giant panda (Ailuropoda melanoleuca), Journal of Natural History, vol. 42, pp.2585-2591, (2008).

DOI: 10.1080/00222930802277624

[28] S. Kumar, K. Tamura, and M. Nei, MEGA3: integrated software for molecular evolutionary genetics analysis and sequence alignment, Briefings in bioinformatics, vol. 5, p.150, (2004).

DOI: 10.1093/bib/5.2.150