›› 2010, Vol. 37 ›› Issue (5): 85-89.

• 生物技术 • Previous Articles     Next Articles

Stably Expressing Research of Interferongamma (IFNγ) from Cervus Nippon in Different Eukaryotic Cell Lines

LIU Ying1,2, WANG Bing1,2, YU Qinglong1,2, XIONG Jiajun3, YANG Liguo3, CHEN Huanchun1,2, GUO Aizhen1,2   

  1. （1.The State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, China;2.College of Veterinary Medicine, Hubei Provincial Key Laboratory of Preventive Veterinary Medicine, Huazhong AgriculturalUniversity, Wuhan 430070, China; 3.College of Animal Science, Huazhong Agricultural University, Wuhan 430070, China）
  • Received:1900-01-01 Revised:1900-01-01 Online:2010-05-20 Published:2010-05-20

Abstract: Total RNA was isolated from Cervus Nippon peripheral blood lymphocytes, which were stimulated with PHA. Then the Cervus Nippon mature IFNγ gene was amplified by reverse transcription chain reaction. The results indicated that the cloned gene was mature Cervus Nippon IFNγ gene, which had the identities of 100% with the CerIFNγ gene published in the GenBank, subcloned into the eukaryotic expression plasmid pCIneo vector which had the CMV enhancer. The recombinant pCIneoCerIFNγ plasmids were transfected into BHK21 cell and MDBK cell by calcium phosphate. The stably expression of transfected cell lines were screened by the G418 resistance. Detected by Western blotting, to determine the relative molecular expression products were 23, 20, 16 ku, it is consistent with the forecast size. This research results lay the foundation for further develop of the Cervus Nippon biological preparations class treatment.

Key words: Cervus Nippon; interferongamma; cell lines; eukaryotic expression

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