Rapid communication: nucleotide sequence of red seabream, Pagrus major, beta-actin cDNA.

Name of the Sequence. Red seabream β-actin cDNA. Genus and Species. Pagrus major. Origin of Clone. Total RNA was isolated from ovary of red seabream. Poly(A)+ RNA was purified using an Oligotex-dT30 (Super) mRNA purification kit (Takara, Japan). Double-stranded complementary DNA synthesis was carried out using a TimeSaver cDNA synthesis kit (Pharmacia Biotech, Uppsala, Sweden). The cDNA library was constructed with a Lamda gt10/EcoRI/ CIAP-treated vector (Stratagene, La Jolla, CA). To target β-actin cDNA from red seabream, primers for PCR were synthesized referring to DNA nucleotide sequences reported for medaka β-actin (Takagiri et al., 1997). The PCR primers used to screen the red seabream ovary cDNA library were ACTACCTCATGAAG ATCCTG for the forward primer and TTGCTGATCCA CATCTGCTG for the reverse primer. Reverse transcriptase-PCR amplified a 517-bp band from red seabream cDNA synthesized from the ovary RNA. The PCR product as a probe was labeled with [α-32P]dCTP (Oligolabelling kit, Pharmacia Biotech) to screen the cDNA. The positive clones containing the insert were digested with Sau3AI, SacI, and ScaI and then subcloned into pBlue Script SK+. Both strands were sequenced with an ALFexpress DNA Sequencer System (Pharmacia Biotech). Comparison with Related Sequences. The deduced amino acid sequence of red seabream β-actin showed 96.3% identity to both human (Ponte et al., 1984) and medaka (Takagiri et al., 1997) and 95.7% identity to gilthead seabream β-actin (Santos et al., 1997).