Characterization and classification of Bo4 as a cluster G mycobacteriophage that can infect and lyse M. tuberculosis
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ORIGINAL PAPER
Characterization and classification of Bo4 as a cluster G mycobacteriophage that can infect and lyse M. tuberculosis Yiling Gan · Tingting Wu · Ping Liu · Shuliang Guo
Received: 21 November 2013 / Revised: 19 January 2014 / Accepted: 21 January 2014 / Published online: 7 February 2014 © Springer-Verlag Berlin Heidelberg 2014
Abstract Mycobacteriophage therapy is a potential alternative treatment for Mycobacterium tuberculosis infection. Here, we further characterized a mycobacteriophage, Bo4, and evaluated its ability to infect and kill M. tuberculosis. We first found that Bo4 can infect M. tuberculosis and Mycobacterium smegmatis. The observed clear plaques created by Bo4 infection indicated that Bo4 might be a lytic phage able to lyse mycobacterial strains, which was confirmed by phage antimicrobial activity. Bo4 formed clear zones in a medium with pH values of 7.4 or 5.0, suggesting the possibility that Bo4 could lyse mycobacteria, such as M. tuberculosis, in blood as well as in lysosomal macrophages. Further investigation into the Bo4 genome revealed that Bo4 had a dsDNA genome. Moreover, Bo4 contained ~39,318 bp comprised of 66.76 % G+C content. Complete genome sequencing showed high nucleotide identity with cluster G mycobacteriophages, thus classifying Bo4 as a
Communicated by Erko Stackebrandt. Y. Gan · S. Guo (*) Department of Respiratory Medicine, The First Affiliated Hospital of Chongqing Medical University, Chongqing 1st Youyi Road, Chongqing 400016, China e-mail: [email protected]; [email protected] Y. Gan e-mail: [email protected] T. Wu Department of Respiratory Medicine, The Third People’s Hospital of Chengdu City, Chengdu, China e-mail: [email protected] P. Liu Department of Respiratory Medicine, The People’s Hospital of Changshou District, Chongqing, China e-mail: [email protected]
member of the cluster G family. Additionally, annotation of the Bo4 genome indicated that it was a lytic bacteriophage and did not contain any harmful genes that increased mycobacterial virulence or decreased human immunity. Overall, the results of investigation indicate that the Bo4 possesses the potential to destroy M. tuberculosis, making it a potentially useful tool for diagnosing and treating tuberculosis. Keywords Lytic mycobacteriophage · Biological characteristics · Genomic analysis
Introduction Mycobacterium tuberculosis is certainly one of the most successful and sophisticated pathogens to infect humans worldwide. Like other infectious diseases, an ever-increasing number of drug-resistant M. tuberculosis strains frequently arise, but the development of new and effective anti-tuberculosis (TB) drugs has been difficult (Tomioka and Namba 2006). Bacteriophages that are capable of lysing their bacterial hosts (O’Flaherty et al. 2009) possess several characteristics that may make them ideal antimicrobial candidates, including high specificity, rapid self-proliferation, and wide availability. Therapy using mycobacteriophages, which specifically infect mycobacteria, has become a highlig
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