TY - JOUR
T1 - Characterization of temperate phage Che12 and construction of a new tool for diagnosis of tuberculosis
AU - Kumar, Vanaja
AU - Loganathan, Prabakaran
AU - Sivaramakrishnan, Gomathi
AU - Kriakov, Jordan
AU - Dusthakeer, Azger
AU - Subramanyam, Balaji
AU - Chan, John
AU - Jacobs, William R.
AU - Paranji Rama, Narayanan
N1 - Funding Information:
VK, LP, NSG and AD each acknowledge support from the AIDS International Training and Research Program (NIH D43-TW01403) of the Albert Einstein College of Medicine (Program Director: Vinayaka Prasad).
Funding Information:
SB acknowledges the support by the World Health Organization with funds from the United States Agency for International Development through South East Asia Regional Office, New Delhi, India.
PY - 2008/11
Y1 - 2008/11
N2 - A temperate phage, Che12, able to infect Mycobacterium tuberculosis, was isolated from soil samples taken from tuberculosis sanatorium area in Chennai, India. The plaque morphology of this phage showed varying grades of turbidity on lawns of M. tuberculosis. The temperate nature of Che12 was established by super infection immunity. Phage integration into the host genomic DNA was confirmed by Southern hybridization using Che12 DNA as a probe. PCR amplification and sequencing of a part of the integrated phage genome in a M. tuberculosis lysogen also confirmed the temperate nature of Che12. The morphology of the phage particles was observed by electron microscopy, revealing similarities to other mycobacteriophages like L5, D29 and TM4. A luciferase reporter phage, phAETRC16, was constructed by cloning firefly luciferase gene into Che12. Infection of viable M. tuberculosis cells by phAETRC16 resulted in expression of luciferase leading to sustained light output. Che12, a true temperate phage infecting M. tuberculosis, is thus ideally suited for developing a diagnostic tool facilitating rapid diagnosis of M. tuberculosis.
AB - A temperate phage, Che12, able to infect Mycobacterium tuberculosis, was isolated from soil samples taken from tuberculosis sanatorium area in Chennai, India. The plaque morphology of this phage showed varying grades of turbidity on lawns of M. tuberculosis. The temperate nature of Che12 was established by super infection immunity. Phage integration into the host genomic DNA was confirmed by Southern hybridization using Che12 DNA as a probe. PCR amplification and sequencing of a part of the integrated phage genome in a M. tuberculosis lysogen also confirmed the temperate nature of Che12. The morphology of the phage particles was observed by electron microscopy, revealing similarities to other mycobacteriophages like L5, D29 and TM4. A luciferase reporter phage, phAETRC16, was constructed by cloning firefly luciferase gene into Che12. Infection of viable M. tuberculosis cells by phAETRC16 resulted in expression of luciferase leading to sustained light output. Che12, a true temperate phage infecting M. tuberculosis, is thus ideally suited for developing a diagnostic tool facilitating rapid diagnosis of M. tuberculosis.
KW - Luciferase reporter phage
KW - M. tuberculosis
KW - Temperate
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U2 - 10.1016/j.tube.2008.02.007
DO - 10.1016/j.tube.2008.02.007
M3 - Article
C2 - 18511339
AN - SCOPUS:54849425143
SN - 1472-9792
VL - 88
SP - 616
EP - 623
JO - Tuberculosis
JF - Tuberculosis
IS - 6
ER -
