RESUMO
We have determined the complete cDNA sequences of fresh water rod opsin gene (fwo) and deep-sea rod opsin gene (dso) from Japanese eel Anguilla japonica. The cDNA clones of fwo and dso consisted of 1437 and 1497 nucleotides, respectively. The predicted opsins of both genes consisted of 352 amino acid residues. Southern blot and PCR analyses of genomic DNA indicated that the Japanese eel genome contains only one fwo and one dso and they are intronless. Quantitative RT-PCR analyses revealed that the expression of fwo decreases with sexual maturation while that of dso increases.
Assuntos
Anguilla/genética , Regulação da Expressão Gênica no Desenvolvimento/genética , Opsinas de Bastonetes/genética , Animais , Southern Blotting , Clonagem Molecular , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Maturidade Sexual/genéticaRESUMO
A cDNA clone of rainbow trout (Oncorhynchus mykiss) transferrin was obtained from a liver cDNA library. The 2537-bp cDNA sequence contained an open reading frame encoding 691 amino acids and the 5' and 3' noncoding regions. The amino acid sequences at the iron-binding sites and the two N-linked glycosylation sites, and the cysteine residues were consistent with known, conserved vertebrate transferrin cDNA sequences. Single N-linked glycosylation sites existed on the N- and C-lobe. The deduced amino acid sequence of the rainbow trout transferrin cDNA had 92.9% identities with transferrin of coho salmon (Oncorhynchus kisutch); 85%, Atlantic salmon (Salmo salar); 67.3%, medaka (Oryzias latipes); 61.3% Atlantic cod (Gadus morhua); and 59.7%, Japanese flounder (Paralichthys olivaceus). The long and accurate polymerase chain reaction (LA-PCR) was used to amplify approximately 6.5 kb of the transferrin gene from rainbow trout genomic DNA. Restriction fragment length polymorphisms (RFLPs) of the LA-PCR products revealed three digestion patterns in 22 samples.
Assuntos
Oncorhynchus mykiss/genética , Polimorfismo Genético , Transferrina/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Clonagem Molecular , Sequência Conservada , Cisteína , Primers do DNA , Glicosilação , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , Alinhamento de Sequência , Homologia de Sequência de Aminoácidos , Transferrina/biossíntese , Transferrina/química , VertebradosRESUMO
Transferrin is a monomeric iron-binding glycoprotein. To investigate the structure of the transferrin gene in teleosts, we prepared a genomic DNA library from the medaka (Oryzias latipes). The medaka transferrin gene was isolated, and its nucleotide sequence was determined. The full length of this gene is approximately 8.5 kb, and it is organized into 17 exons separated by 16 introns. The exons are similar in size to those in the genes for human transferrin, chicken ovotransferrin, and mouse and bovine lactoferrin. However, the introns are smaller than those previously reported in the transferrin family of genes. The 5'-flanking region of the medaka transferrin gene contains an estrogen-responsive element, glucocorticoid-responsive elements, an iron-responsive element, metal-responsive elements, and Sp1 binding sites. The transcription start point is located 35 bp upstream of the start codon.