牛干扰素-γ基因的克隆、序列分析及表达研究

Abstract

Abstract: In order to build more effective use of interferon-gamma (IFN-γ) on the diagnosis and control of diseases like bovine tuberculosis,the cDNA of bovine interferon-γ (BoIFN-γ) gene was prepared from PHA-stimulated Yunnan cattle periphetal blood lymphocyte cell by RT-PCR. The BoIFN-γ gene were identified by sequencing and BLAST. Then,the BoIFN-γ was inserted into the expressing plasmid pET-28a for expression in E. coli BL21 (DE3). The recombinant bacteria were induced with IPTG. Sequence analysis showed the homology of BoIFN-γ gene between Yunnan and Holstein cows was 99.60%. The molecular size of IFN-γ expressed was 23 ku with SDS-PAGE electrophoresis detection,which was consistent with the forecast. The analysis results of ELISA Kits showed the biological activity of IFN-γ expressed was obviously,the content in bacteria was 244 ng/mL. The experiment lays a foundation for the further research and application of bovine IFN-γ.

Key words: bovine interferon-gamma; sequence analysis; prokaryotic expression; biological activity

CLC Number:

YANG Jie, ZHANG Shu-guang, LI Chang-qing, LIU Shi-gao, CHAI Jun, LIU Xu-chuan, SHI Xian-wei, ZHANG Yi-fang. Cloning,Sequence Analysis and Expression of Bovine Interferon-gamma Gene[J]. , 2014, 41(10): 1-6.

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Cloning,Sequence Analysis and Expression of Bovine Interferon-gamma Gene

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