FIG. 2.

(A) Transcription analysis was performed by RT-PCR. Primers A to D, annealing at nucleotides 5299 to 5276, nucleotides 4355 to 4336, nucleotides 3563 to 3546, and nucleotides 2300 to 2280, respectively, were used for RT. PCR of chromosomal DNA was used as a positive control (lanes DNA). The PCR performed on mRNA subjected to an RT reaction is shown in lanes RT; PCR that was performed on mRNA without prior RT reaction (lanes RNA) served as a negative control. Lane M, molecular mass marker (in nucleotides). (B) RT-PCR analysis of the transcription start site. RT reactions were carried out with a primer annealing at nucleotides 2300 to 2280 of the cyl gene cluster. Subsequent PCRs were performed with forward primers annealing upstream (G) or downstream (E and F) of the putative transcription terminator at nucleotides 35 to 55, nucleotides 352 to 372, and nucleotides 301 to 321, respectively. Positive and negative controls are indicated.