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Cloning, expression and bioactivity of chicken receptor activator of NF-κB ligand (chRANKL)

Published online by Cambridge University Press:  03 March 2009

Wang Yan  and
Hou Jia-Fa
Wang Yan
Affiliation:
College of Veterinary Medicine, Nanjing Agricultural University, Nanjing210095, China
Hou Jia-Fa*
Affiliation:
College of Veterinary Medicine, Nanjing Agricultural University, Nanjing210095, China
*
*Corresponding author. E-mail: [email protected]
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Abstract

Using reverse transcription polymerase chain reaction (RT-PCR) and gene splicing by overlap extension (SOE-PCR), the DNA sequence encoding the chicken receptor activator of NF-κB ligand (RANKL) was amplified, cloned into the expressing plasmid pET-32a(+) with His-tag, and highly expressed in Escherichia coli. The purified protein was then added to primary cultures of chicken osteoclasts to observe its bioactivity. The results showed that the size of the PCR product was 1200 bp, which was consistent with the expected size, and the relative molecular weight of the induced protein was 64 kDa. Western blotting indicated that induced protein could react with anti-His antibody. It was also found that the induced protein can stimulate mature chicken osteoclasts to resorb bone.

Keywords

chRANKLgene cloningprokaryotic expression
Type
Research Papers
Information
Chinese Journal of Agricultural Biotechnology , Volume 5 , Issue 3 , December 2008 , pp. 205 - 209
Copyright
Copyright © China Agricultural University 2008

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Footnotes

First published in Journal of Agricultural Biotechnology 2008, 16(1): 32–36

References

Blair, JM, Zhou, H, Seibel, MJ and Dunstan, CR (2006) Mechanisms of disease: role of OPG, RANKL and RANK in the pathophysiology of skeletal metastasis. Nature Clinical Practice Oncology 3: 41–49.Google Scholar
Burgess, TL, Qian, YX, Kaufman, S, et al. (1999) The ligand for osteoprotegerin (OPGL) directly activates mature osteoclasts. Cell Biology 145(3): 527–538.CrossRefGoogle ScholarPubMed
Fujikawa, Y, Sabokbar, A, Neale, SD, Itonaga, I, Torisu, T and Athanasou, NA (2001) The effect of macrophage-colony stimulating factor and other humoral factors (interleukin-1, -3, -6 and -11, tumor necrosis factor-alpha, and granulocyte macrophage-colony stimulating factor) on human osteoclast formation from circulating cells. Bone 28(3): 261–267.Google Scholar
Lacey, DL, Timms, E, Tan, HL, et al. (1998) Osteoprotegerin ligand is a cytokine that regulates osteoclast differentiation and activation. Cell 93(2): 165–176.Google Scholar
Matsuzaki, K, Udagawa, N, Takahashi, N, et al. (1998) Osteoclast differentiation factor (ODF) induces osteoclast-like cell formation in human peripheral blood mononuclear cell cultures. Biochemical and Biophysical Research Communications 246: 199–204.CrossRefGoogle ScholarPubMed
Quinn, JM, Neale, S, Fujikawa, Y, McGee, JO and Athanasou, NA (1998) Human osteoclast formation from blood monocytes, peritoneal macrophages, and bone marrow cells. Calcified Tissue International 62(6): 527–531.CrossRefGoogle Scholar
Takahashi, N, Udagawa, N and Suda, T (1999) A new member of tumor necrosis factor ligand family, ODF/OPGL/TRANCE/RANKL, regulates osteoclast differentiation and function. Biochemical and Biophysical Research Communications 256: 449–455.Google Scholar
Whitehead, CC (2004) Skeletal disorders in laying hens: the problems of osteoporosis and bone fractures. Poultry Science Symposium 27: 259–278.Google Scholar
Wong, BR, Josien, R, Lee, SY, et al. (1997) TRANCE (tumor necrosis factor [TNF]-related activation-induced cytokine), a new TNF family member predominantly expressed in T cells, is a dendritic cell-specific survival factor. Journal of Experimental Medicine 186(12): 2075–2080.Google Scholar
Yao, J, Qian, C-Z and Hou, J-F (2005) Isolation, culture and identification of osteoclasts from the embryonic chicken. Journal of Nanjing Agricultural University 8(3): 88–91.Google Scholar
Yao, J, Zhang, J-F and Hou, J-F (2007) Effects of Ipriflavone on caged layer bone metabolism in vitro and vivo. Poultry Science 86(3): 503–507.Google Scholar
Yasuda, H, Shima, N, Nakagawa, N, et al. (1998) Osteoclast differentiation factor is a ligand for osteoprotegerin/osteoclastogenesis-inhibitory factor and is identical to TRANCE/RANKL. Proceedings of the National Academy of Sciences, USA 95(7): 3597–3602.Google Scholar