Bacterial transcript imaging by hybridization of total RNA to oligonucleotide arrays.
We have used high-density oligonucleotide probe arrays (chips) for bacterial transcript imaging. We designed a chip containing probes representing 106 Hemophilus influenzae genes and 100 Streptococcus pneumoniae genes. The apparent lack of polyadenylated transcripts excludes enrichment of mRNA by affinity purification and we thus used total, chemically biotinylated RNA as hybridization probe. We show that hybridization of Streptococcus RNA to a chip allows simultaneous quantification of the transcript levels. The sensitivity was found to be in the range of one to five transcripts per cell. The quantitative chip results were in good agreement with conventional Northern blot analysis of selected genes. This technology allows simultaneous and quantitative measurement of the transcriptional activity of entire bacterial genomes on a single oligonucleotide probe array.
de Saizieu
A
A
Pharma Division, F. Hoffmann-La Roche Ltd., Basel, Switzerland.
Certa
U
U
Warrington
J
J
Gray
C
C
Keck
W
W
Mous
J
J
D019921:Biotinylation; D015152:Blotting, Northern; D005456:Fluorescent Dyes; D016680:Genome, Bacterial; D006193:Haemophilus influenzae / Q000235:genetics*; D009693:Nucleic Acid Hybridization; D015345:Oligonucleotide Probes; D012329:RNA, Bacterial; D013296:Streptococcus pneumoniae / Q000235:genetics*; D014158:Transcription, Genetic / Q000235:genetics*
D005456:Fluorescent Dyes; D015345:Oligonucleotide Probes; D012329:RNA, Bacterial