Characterization of prokaryotic mRNAs by RT-PCR
Runar Ra, S.; Saris, P.E.
Biotechniques 18(5): 792-795
1995
ISSN/ISBN: 0736-6205 PMID: 7542457 Document Number: 451941
The use of polymerase chain reaction (PCR) techniques for the analysis of bacterial gene expression at the RNA level was investigated using expression of nisin genes in Lactococcus lactis as a model system; in particular, the possible co-transcription of nisZ and nisB genes as polycistronic mRNA was investigated using reverse transcription (RT) and subsequent PCR amplification of the resulting cDNA. The RT-PCR technique was successful in amplifying the expected 630-bp RNA sequence, and digestion with SacI yielded the predicted 500- and 130-bp fragments. It is concluded that a polycistronic nisZB transcript is produced, and this supports the hypothesis that the inverted repeat between nisZ and nisB functions as a terminator and regulates the amount of transcripts of the biosynthetic genes downstream of the structural gene of the nisin gene cluster. The general use of RT-PCR techniques for prokaryotic RNA studies is discussed.