Flourescence-based isolation of differentially induced genes

Details Flourescence-based isolation of differentially induced genes Flourescence-based isolation of differentially induced genes

1997-09-10

1999-11-30

Eisenchenk, Frank C.

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving nucleic acid

435 691, 435189, 435 697, 4352523, 536 231, 536 234, 536 235, 536 221, C12Q 168, C12P 2106, C12P 2104, C12N 902

Patent

active

059940778

ABSTRACT:
Regulatory elements (e.g. promoters) activated by a stimulus are isolated by a FACS-based method. Preferably, a library of random fragments representative of a target (e.g. bacterial) genome are cloned in front of a promoterless gfp (green fluorescent protein) sequence in a plasmid, and inserted into target cells. The resulting target cell mixture is sorted according to GFP levels in the presence and the absence of the stimulus. Suitable stimuli include compounds of interest (e.g. drugs), environmental factors (e.g. extracellular acidity), and complex stimuli such as in vivo environments of hosts infected by the target cells. The method allows identifying pathogen genes which are selectively expressed during infection.

REFERENCES:
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Heithoff, D. et al., Bacterial infection as assessed by in vivo gene expression, Proc. Natl. Acad. Sci. USA, vol. 94, pp. 934-939, Feb. 1997.
Mahan, M. et al., Antibiotic-based selection for bacterial genes that are specifically induced during infection in host, Proc. Natl. Acad. Sci. USA, vol. 92, pp. 669-673, Jan. 1995.
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Mahan, et al. : Antibiotic-based selection for bacterial genes...: Proc. Natl. Acad. Sci.: vol. 92: pp. 669-673, Jan. 1995.

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