title: MOLECULAR CLONING, SEQUENCING, EXPRESSION AND BIOLOGICAL ACTIVITY OF GIANT PANDA (AILUROPODA MELANOLEUCA) INTERFERON-GAMMA.
abstract: The giant panda (Ailuropoda melanoleuca) is an endangered species and indigenous to China. Interferon-gamma (IFN-γ) is the only member of type □ IFN and is vital for the regulation of host adapted immunity and inflammatory response. Little is known aboutthe FN-γ gene and its roles in giant panda.In this study, IFN-γ gene of Qinling giant panda was amplified from total blood RNA by RT-CPR, cloned, sequenced and analysed. The open reading frame (ORF) of Qinling giant panda IFN-γ encodes 152 amino acidsand is highly similar to Sichuan giant panda with an identity of 99.3% in cDNA sequence. The IFN-γ cDNA sequence was ligated to the pET32a vector and transformed into E. coli BL21 competent cells. Expression of recombinant IFN-γ protein of Qinling giant panda in E. coli was confirmed by SDS-PAGE and Western blot analysis. Biological activity assay indicated that the recombinant IFN-γ protein at the concentration of 4-10 µg/ml activated the giant panda peripheral blood lymphocytes,while at 12 µg/mlinhibited. the activation of the lymphocytes.These findings provide insights into the evolution of giant panda IFN-γ and information regarding amino acid residues essential for their biological activity.
authors: a College of Veterinary Medicine, Northwest Agriculture and Forestry University , Yangling , Shaanxi Province , P. R. China.
authors: Hui
authors: H
authors: Zhu
authors: Wen-Xiu
authors: WX
authors: Wang
authors: Bao-Qin
authors: BQ
authors: Wang
authors: Xiao-Fu
authors: XF
authors: Zhu
authors: Xu-Jin
authors: XJ
authors: Wu
authors: Qing-Yi
authors: QY
authors: Ma
authors: De-Kun
authors: DK
authors: Chen
keywords: IFN-γ; IFN-γ biological activity; IFN-γ cloning; IFN-γ expression; giant panda (ailuropoda melanoleuca)