›› 2011, Vol. 38 ›› Issue (12): 96-99.

• 生物技术 • Previous Articles     Next Articles

Construction of Eukaryotic Expression Vector and Sequence Analysis of Antimicrobial Peptide Gene Shiva 1a

GONG Xiao-wei, ZHENG Fu-ying, LIN Guo-zhen, CAO Xiao-an, WANG Guang-hua, ZHOU Ji-zhang, CAI Xue-peng    

  1. State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Public Health of the Ministry of Agriculture,Lanzhou Veterinary Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, China
  • Received:2011-04-16 Revised:1900-01-01 Online:2011-12-20 Published:2011-12-20

Abstract: To explore the effect mechanism of cecropin-class lytic peptide at early stage of infection. The mat peptide of Shiva 1a was amplified by overlap extension PCR and the C-terminus contained 6×His-marker. The gene sequence were recombinant with eukaryotic expression vector pIRES2-EGFP. After being identified by restriction enzyme digestion and sequencing, the recombinant plasmid pIRES2-EGFP-Shiva 1a was transfected into CHO-K1 cells by liposomes. The expression of the the recombinant plasmid pIRES2-EGFP-Shiva 1a was observed by fluorescence microscope. At the same time, the secondary structure and 3D structure were predicted by bioinformatics tools. The results lay the foundation in research of antimicrobial activities and applications of the antimicrobial peptide Shiva 1a in breeding for disease resistance of animals.

Key words: antimicrobial peptide; Shiva 1a; eukaryotic expression; sequence analysis

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