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Infection and Immunity, January 2000, p. 387-390, Vol. 68, No. 1
Emory University School of Medicine and
Centers for Disease Control and Prevention, Atlanta, Georgia
Received 10 May 1999/Returned for modification 5 July 1999/Accepted 5 October 1999
A coinfection assay was developed to examine Mycobacterium
tuberculosis genes suspected to be involved in resistance to
killing by human macrophages. THP-1 macrophages were infected with a
mixture of equal numbers of recombinant Mycobacterium
smegmatis LR222 bacteria expressing an M. tuberculosis gene and wild-type M. smegmatis LR222
bacteria expressing the xylE gene. At various times after infection, the infected macrophages were lysed and the bacteria were
plated. The resulting colonies were sprayed with catechol to determine
the number of recombinant colonies and the number of
xylE-expressing colonies. M. smegmatis bacteria
expressing the M. tuberculosis glutamine synthetase A
(glnA) gene or open reading frame Rv2962c or
Rv2958c demonstrated significantly increased survival rates
in THP-1 macrophages relative to those of xylE-expressing bacteria. M. smegmatis bacteria expressing M. tuberculosis genes for phospholipase C (plcA and
plcB) or for high temperature requirement A
(htrA) did not.
0019-9567/0/$04.00+0
Evaluation of Mycobacterium tuberculosis
Genes Involved in Resistance to Killing by Human Macrophages
and
*
Corresponding author. Mailing address: Centers for
Disease Control and Prevention, Mailstop G35, 1600 Clifton Rd.,
Atlanta, GA 30329. Phone: (404) 639-3601. Fax: (404) 639-1287. E-mail: tms1{at}cdc.gov.
Present address: Department of Internal Medicine, University of
Michigan School of Medicine, Ann Arbor, Mich.
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