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| Expression of the fusion gene encoding bovine antimicrobial peptides Bac7-Bac5-β defense in insect rod-shaped virus system, and antimicrobial activity of the fusion protein |
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Abstract In the present study, based on the gene sequences encoding mature bovine antimicrobial peptides bac7、bac5 and - defense（LAP） as registered in Genbank, the fragment of the fusion gene Bac7-Bac5-βdefense was synthesized and cloned into the pFASTHTb vector of BAC-TO-BACTM recombinant baculovirus expression system to construct a recombinant expression vector, The plasmid pFASTBac-B7-B5- was then transformed into DH10BACTM competent cell. High molecular weight DNA, the recombinant bacmid, was prepared from the overnight cultures of selected E. coli DH10BACTM colonies. By transfecting Sf9 cells with the recombinant bacmid the recombinant baculovirus from the superenatant was obtained, which could further infect Sf9 cells obtained and express the target protein. The expression of rB7-B5- was confirmed by 12% SDS-PAGE and accounted for 10.6％ of total bacterial protein,the molecular weight of rB7-B5- was 20kD, a figure consistent with the predicted value. rB7-B5- showed antimicrobial activity in E. coli. The present study provides basis for the research and development of novel antibacterial preparations.
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Received: 10 April 2007
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