RESUMEN
A full-length cDNA clone of a canine carbonic anhydrase VI (CA-VI) was generated from the canine parotid gland by using a reverse transcription-polymerase chain reaction (RT-PCR) technique with degenerate primers designed from conserved regions of the same locus in humans and bovines employing RACE (rapid amplification of cDNA ends) techniques. The cDNA sequence was 1351 base pairs (bp) long and was predicted to encode a 320-amino-acid polypeptide containing a putative signal peptide of 17 amino acids. The deduced amino acid sequence of mature CA-VI showed the highest similarity of 74% to that of human CA-VI. RT-PCR analysis with primers specific to the canine CA-VI demonstrated strong signals in the major salivary glands and weak signals in the minor salivary glands and esophagus of a healthy dog. No CA-VI mRNA was detected in the pancreas, liver or the digestive tract except the esophagus.
Asunto(s)
Anhidrasa Carbónica IV/genética , Perros/genética , Expresión Génica , Animales , Secuencia de Bases , Cartilla de ADN , ADN Complementario/genética , Perfilación de la Expresión Génica , Datos de Secuencia Molecular , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Análisis de Secuencia de ADN , Homología de SecuenciaRESUMEN
Wee1 is one of the genes which regulates the cell cycle. cDNA encoding a Wee1-homolog has been isolated from Japanese silver crucian carp (ginbuna) by using polymerase chain reaction (PCR) techniques. The cDNA obtained from the ovary tissue encoded a protein of 526 amino acids and the deduced amino acid sequence showed high homology (69.9%) with Xenopus laevis Wee1 in the protein kinase domain. A phylogenetic analysis among Wee1 amino acid sequences from various organisms showed that the crucian carp Wee1 was closely related to Xenopus Wee1 and human Wee1B. The vertebrate Wee1s were classified into two types: the meiotic type and the mitotic type. The crucian carp Wee1 described here belonged to the meiotic type. Furthermore, a long-PCR technique allowed us to isolate the wee1 gene of almost full length. The wee1 gene was about 6.3 kbp in length and contained 12 exons, with the open reading frame starting at the second exon.
