TY - JOUR
T1 - Survival of mice infected with Mycobacterium smegmatis containing large DNA fragments from Mycobacterium tuberculosis
AU - Bange, F. C.
AU - Collins, F. M.
AU - Jacobs, W. R.
N1 - Funding Information:
The Institutional Biosafety Committee of the Albert Einstein College of Medicine has reviewed the data presented in this paper and authorized that the M. tuberculosis genomic library described in this paper may be introduced into M. smegmatis in BSL-2 containment facilities. This action has been supported by the Recombinant DNA Advisory Committee of the National Institute of Health.
Funding Information:
We wish to thank John D. McKinney for performing the EMS mutagenesis on M. smegmatis mc2155, George Heckel for helping map the arginine auxotrophs of M. smegmatis, Bing Chen, Elizabeth Fiugeroa, Cynthia Kelley and Angela Howard for technical assistance on the animal experiments, and Mary K. Hondalus and Jordan Kriakov for critical reading of and constructive comments on the manuscript. F.-Ch. Bange was supported by a postdoctoral fellowship of the ‘Infektionsforschung und AIDS – Stipendiumprogramm’ of the German government, and the work was supported by NIH AI26170.
PY - 1999/6
Y1 - 1999/6
N2 - Mycobacterium smegmatis is typically used as a bacterial host for cloning and expressing single genes or genomic libraries of the human pathogen Mycobacterium tuberculosis. To study virulence of M. tuberculosis, we set out to ask the question, whether a genomic library derived from M. tuberculosis H37Rv confers virulence to the non-virulent M. smegmatis. A representative library from the M. tuberculosis H37Rv genome was generated and transformed into wild-type M. smegmatis. Mice were challenged with recombinant clones by intravenous, aerogenic and intranasal infection. We were unable to detect either growth or persistence of recombinant clones in tissues of infected mice; instead, the infection was cleared. Since the concern that virulent traits might be transferred, biosafety regulations often require the handling of these experiments at Biosafety Level 3. However, we failed to find any evidence that the M. tuberculosis library confers virulence when expressed in M. smegmatis. We suggest that the results, presented here, should fundamentally alter the containment requirements for similar experiments in the future.
AB - Mycobacterium smegmatis is typically used as a bacterial host for cloning and expressing single genes or genomic libraries of the human pathogen Mycobacterium tuberculosis. To study virulence of M. tuberculosis, we set out to ask the question, whether a genomic library derived from M. tuberculosis H37Rv confers virulence to the non-virulent M. smegmatis. A representative library from the M. tuberculosis H37Rv genome was generated and transformed into wild-type M. smegmatis. Mice were challenged with recombinant clones by intravenous, aerogenic and intranasal infection. We were unable to detect either growth or persistence of recombinant clones in tissues of infected mice; instead, the infection was cleared. Since the concern that virulent traits might be transferred, biosafety regulations often require the handling of these experiments at Biosafety Level 3. However, we failed to find any evidence that the M. tuberculosis library confers virulence when expressed in M. smegmatis. We suggest that the results, presented here, should fundamentally alter the containment requirements for similar experiments in the future.
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U2 - 10.1054/tuld.1998.0201
DO - 10.1054/tuld.1998.0201
M3 - Article
C2 - 10656115
AN - SCOPUS:0345621667
SN - 0962-8479
VL - 79
SP - 171
EP - 180
JO - Tubercle and Lung Disease
JF - Tubercle and Lung Disease
IS - 3
ER -
