ACTA VETERINARIA ET ZOOTECHNICA SINICA ›› 2009, Vol. 40 ›› Issue (5): 725-729.doi:

• 预防兽医 • Previous Articles     Next Articles

Prokaryotic Expression and Purification of Bioactivity Analysis of Chicken Granulocyte Macrophage Clony Stimulating Factor（GM-CSF）

TAN Bing1, WANG Hong-ning1，2*，ZHANG Yi2,ZHANG An-yun2,FAN Wen-qiao2   

  1. 1. College of Animal Medicine, Sichuan Agricultural University, Yaan 625014, China; 2. Animal Disease Prevention and Food Safety Key Laboratory of Sichuan Province,College of Life Science, Sichuan University, Chengdu 610064, China
  • Received:1900-01-01 Revised:1900-01-01 Online:2009-05-25 Published:2009-05-25

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Abstract: The ChGM-CSF gene without signal peptide was amplified by PCR method and ligated into the expression plasmid pET-32a(+) to construct prokaryotic plasmid p32GM-CSF. ChGM-CSF recombinant protein was expressed by induction with IPTG in E.coli and purified with Ni-NTA column. MTT assay was used to detect bioactivity of rChGM-CSF. Polyclonal antibody was produced by rabbit injected with the purified protein. The result showed that expression plasmid p32GM-CSF was successfully constructed. SDS-PAGE demonstrated that the recombinant protein was expressed in form of inclusion body and was approximately 34 ku. Western Blot analysis showed that it could be specifically recognized by the rabbit sera to chicken GM-CSF. Results of MTT assay confirmed that it can enhance chicken bone marrow cell proliferation obviously. These results demonstrated that the E. coli-derived rChGM-CSF and polyclonal antibody against rChGM-CSF have some bioactivities. Our results would provide foundation for further research of biology characteristics and application of ChGM-CSF.