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Genetic information of mycobacteriophages DNAⅢ and its anti-tuberculosis potential

GAN Yi-ling1, LIU Ping2, WU Ting-ting3, GUO Shu-liang1   

  1. 1.Department of Respiratory and Critical Care Medicine, the First Affiliated Hospital of Chongqing Medical  University, Chongqing 400016, China; 2.Department of Respiratory Medicine, People's Hospital of Changshou District, Chongqing 401220, China; 3.Department of Respiratory Medicine, the Third People's Hospital of Chengdu, Chengdu 610031, China
  • Online:2013-10-28 Published:2013-10-31
  • Supported by:

    National Major Science and Technology Program in “Twelfth Five-year” Plan, 2012ZX10003-009; National Key Clinical Specialty Foundation, 2012-949

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Abstract:

Objective To understand the genetic information of mycobacteriophage DNAⅢ, and explore its anti-tuberculosis potential. Methods Mycobacteriophage DNAⅢ DNA was isolated and purified using lambda DNA extraction protocol. The shotgun sequences were assembled with Phred and PhraD and edited with Consed, then gaps were filled by sequencing PCR fragments. The general characteristics of genome were analysed using EditSeq software of DNAStar software package. The genome was scanned for open reading frames using Glimmer 3.0.  The genetic information of mycobacteriophage DNAⅢ genome was further explored by colinearity analysis and phylogenetic tree construction. The ability of DNAⅢ to lyse M.tuberculosis was investigated in vitro, and the effects of temperature, alcohol, pH values on DNAⅢ survival was determined. Results The genome of DNAⅢ possessed 39 520 bp in size, and was linear double-stranded DNA, with G+C content of 66.83%. DNAⅢ contained 60 predicted protein-coding genes, and did not encode any tRNAs. There was no tandem repeat sequence in the genome of DNAⅢ. DNAⅢ gene 32 encoded integrase, and there was also no gene encoding a repressor. DNAⅢ was highly similar and colinear in nucleotide acid level to Angel and BPs. At three different time points (1 d, 2 d and 3 d), the numbers of M.tuberculosis in DNAⅢ group were significantly smaller than those in control group (P<0.05). DNAⅢ was unstable to temperature, alcohol and pH values. Conclusion DNAⅢ, which belongs to cluster G, is a mycobacteriophage capable of lysing M.tuberculosis.

Key words: mycobacteriophage DNAⅢ, genome, M.tuberculosis, anti-tuberculosis