* Your assessment is very important for improving the workof artificial intelligence, which forms the content of this project
Download Rapid communication: Nucleotide sequence of red seabream
Survey
Document related concepts
Community fingerprinting wikipedia , lookup
Ancestral sequence reconstruction wikipedia , lookup
Fatty acid synthesis wikipedia , lookup
Bisulfite sequencing wikipedia , lookup
Non-coding DNA wikipedia , lookup
Real-time polymerase chain reaction wikipedia , lookup
Peptide synthesis wikipedia , lookup
Deoxyribozyme wikipedia , lookup
Protein structure prediction wikipedia , lookup
Nucleic acid analogue wikipedia , lookup
Point mutation wikipedia , lookup
Artificial gene synthesis wikipedia , lookup
Amino acid synthesis wikipedia , lookup
Biochemistry wikipedia , lookup
Transcript
Rapid communication: Nucleotide sequence of red seabream, Pagrus major, β-actin cDNA C. Y. Choi1 and F. Takashima Department of Aquatic Biosciences, Tokyo University of Fisheries, Konan, Minato-ku, Tokyo 108-8477, Japan 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). 1 Present address for correspondence: Dr. Cheol Young Choi, Sesoko Station, Tropical Biosphere Research Center, University of the Ryukyus, 3422 Sesoko, Motobu, Okinawa 905-0227, Japan (phone: +81 980 47 6215; fax: +81 980 47 4919; E-mail: [email protected]). Received January 20, 2000. Accepted July 20, 2000. 2000 American Society of Animal Science. All rights reserved. Sequence Data. The red seabream β-actin cDNA clone contained 1,521 nucleotides (nt), including the open reading frame that encoded 375 amino acids. The stop codon was followed by a 3′ untranslated region of 342 nt, including the polyadenylation signal, AATAAA. DDBJ/EMBL/GenBank Accession Number. AB036756. Comments. From the total of 375 amino acids, the human β-actin amino acid sequences differed by 7 amino acids and 14 amino acids from gilthead seabream and red seabream β-actin amino acid sequences, respectively (Figure 1). The amino acid sequences of red seabream β-actin differed from those of β-actin of gilthead seabream, which belongs to the same family as red seabream, by 16 amino acids out of 375. Comparison of the deduced amino acid sequence of the red seabream β-actin gene was found to be highly conserved in relation to other known actins. However, the red seabream β-actin isoform has a higher homology to human β-actin than β-actin of gilthead seabream, which belongs to the same family as red seabream. Therefore, the possibility that red seabream β-actin and gilthead seabream βactin have different isoforms must be considered. Literature Cited Katagiri, T., I. Hirono, and T. Aoki. 1997. Identification of a cDNA for Medaka cytoskeletal β-actin and construction for the reverse transcriptase-polymerase chain reaction (RT-PCR) primers. Fisheries Sci. (Tokyo) 63:73–76. Ponte, P., S.-Y. Ng, J. Engel, P. Gunning, and L. Kedes. 1984. Evolutionary conservation in the untranslated regions of actin mRNAs: DNA sequence of human beta-actin cDNA. Nucleic Acids Res. 12:1687–1696. Santos, C. R. A., D. M. Power, P. Kille, L. Llewellyn, V. Ramsurn, T. Wigham, and G. E. Sweeney. 1997. Cloning and sequencing of full-length seabream (Sparus aurata) β-actin cDNA. Comp. Biochem. Physiol. 117B:185–189. Key Words: Actin, Complementary DNA, DNA Cloning, Red Seabream 2990 J. Anim. Sci. 2000. 78:2990–2991 Rapid Communication 2991 Figure 1. The amino acid sequences were extracted from the DDBJ/EMBL/GenBank sequence databases. Comparison of the amino acid sequences encoded by the red seabream β-actin (RsACT) (this paper, AB036756) with gilthead seabream (GsACT) (X89920), medaka (MeACT) (D89627), and human β-actin (HuACT) (84144061). The amino acid sequences were optimally aligned so as to match identical residues, which are indicated by asterisks.