Expression and Antibacterial Activity Verification of Endolysin in Prophages of Klebsiella pneumoniae from Cattle
[Objective]The purpose of this experiment was to predict and analyze the prophage carried by Klebsiella pneumoniae and its carrying endolysin gene,explore the antibacterial activity of endolysin in vitro,and provide a new treatment for Klebsiae pneumoniae infection.[Method]Using 134 strains of bovine-origin Klebsiella pneumoniae as the research subjects,the prophages in bovine-origin Klebsiella pneumoniae were predicted through the PhiSpy software.Functional annotation and identification of endolysin were performed on the predicted prophages.The recombinant protein was induced and purified by constructing endolysin prokaryotic expression vector,and its lytic activity against Klebsiella pneumoniae NMG10,GS4 and ATCC 13883 was verified in vitro.[Result]The prophage sequence carriage rate in the genomes of 134 strains of bovine-origin Klebsiella pneumoniae was 98.5％,with a total of 414 predicted prophage sequences.Among them,112 belonged to Klebsiella pneumoniae phages,while the other were either phages or phage components of other bacteria.The proportion of all prophages carried by each bovine-origin Klebsiella pneumoniae genome was approximately 1.4％.A total of 44 prophage genes encoding endolysin were found in the genome of 112 prophages of Klebsiella pneumoniae,which was named LyKb.Endolysins LyKb exhibited both lytic and antibacterial activities against Klebsiella pneumoniae strains NMG10,GS4 and ATCC 13883.Among these,LyKb showed the strongest lytic and antibacterial activities against Klebsiella pneumoniae NMG10,followed by Klebsiella pneumoniae GS4 and ATCC 13883.[Conclusion]134 strains of bovine-origin Klebsiella pneumoniae commonly carried prophages,and some of the prophage genomes contained genes encoding endolysins.The endolysins of Klebsiella pneumoniae prophages had antibacterial activity against various strains of Klebsiella pneumoniae.
万方数据
维普
