Measurement of transcriptional activity in pathogenic bacteria recovered directly from infected host tissue.
In order to understand the genetic regulation of bacterial genes whose products are important for pathogenesis, one needs to measure the expression of the genes during the infection process. We have devised a method to measure the transcriptional activity of such genes from bacteria recovered directly from infected host tissue. Starting with bacterial strains containing lacZ transcriptional fusions to the genes of interest, animals can be infected, with subsequent isolation of infected host tissue. Here we describe the separation of bacterial cells away from a particular host tissue and the subsequent measurement of the activity of beta-galactosidase, the product of the lacZ gene, in the bacterial cells. This assay is sensitive enough to compensate for the potentially low number of bacteria recovered from the infection site.
Harvard Medical School, Boston, MA.
J M
JM
Slauch
M J
MJ
Mahan
J J
JJ
Mekalanos
D000818:Animals; D001419:Bacteria / Q000235:genetics* / Q000302:isolation & purification; D001424:Bacterial Infections / Q000382:microbiology*; D005452:Fluoresceins / Q000378:metabolism; D005697:Galactosides / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D011993:Recombinant Fusion Proteins; D012486:Salmonella typhimurium / Q000235:genetics / Q000302:isolation & purification; D013050:Spectrometry, Fluorescence; D013154:Spleen / Q000382:microbiology; D014158:Transcription, Genetic; D001616:beta-Galactosidase / Q000032:analysis / Q000235:genetics / Q000378:metabolism
D005452:Fluoresceins; D005697:Galactosides; D011993:Recombinant Fusion Proteins; C038848:fluorescein-digalactoside; D001616:beta-Galactosidase