FIG. 4.

RB6K 5′ upstream sequence. (A) The sequence shown corresponds to nucleotides 102787 to 103454 from GenBank accession no. AC004826. Indicated are transcription initiation sites (horizontal arrows), the longest cDNA isolated by 5′-RACE (vertical arrow), the open reading frame (in boldface type and underlined), and the CDE-CHR element (box). (B) Alignment of CDE-CHR element sequences from cell cycle-regulated genes. The consensus sequence as originally described for cdc2, cyclin A, and cdc25C (35) is shown in boxes. (C) Primer extension mapping of the transcription initiation site. Sequence analysis and primer extention assay were performed using a Cy5-labeled oligonucleotide primer. The reaction products were analyzed on an ALF-express sequence analyzer. Lanes A, C, G, and T contain sequence ladder. Lanes − and + contain primer extension reaction mixtures using 5 μg of total RNA isolated from EC-RF24 cells without (−) and with (+) reverse transcriptase added.