RESUMEN
Full-length cDNA that encodes feline α1-microglobulin (Feα1m)-bikunin was obtained from a feline liver and cloned using an oligo-capping method. The Feα1m-bikunin cDNA was found to contain 1284 nucleotides, and Feα1m was found to include an open reading frame encoding a polypeptide of 201 amino acids. The deduced amino acid sequence of Feα1m showed varying amino acid identity when compared with the published sequences of the related α1-m of other species, ranging from 71.1 to 82.1%. Feα1m mRNA expression was confirmed by reverse transcription polymerase chain reaction (RT-PCR) and real-time PCR analysis in the cerebrum, cerebellum, lung, heart, liver, spleen, pancreas, kidney, adrenal gland, and testicle. The highest Feα1m mRNA level was found in the liver.
Asunto(s)
alfa-Globulinas/genética , alfa-Globulinas/biosíntesis , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Gatos/genética , Clonación Molecular , Hígado/metabolismo , Sistemas de Lectura Abierta/genética , ARN Mensajero/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/veterinaria , Alineación de SecuenciaRESUMEN
Cystatin C is a member of the cystatin superfamily of low-molecular-weight proteins that inhibit the activity of cysteine protease. The full-length cDNA that encodes feline cystatin C (FeCysC) has been cloned from feline white blood cells by the oligo-capping method. The cDNA consists of 796 nucleotides and includes an open reading frame encoding a polypeptide of 146 amino acids. Next, we developed several mouse anti-FeCysC monoclonal antibodies (mAbs). The recombinant FeCysC (rFeCysC) was expressed in Escherichia coli (E. coli) BL21 and used as an antigen to immunise mice. The reactivity of the mAbs to native FeCysC was examined by western blot analysis against the urinary protein from cats with chronic kidney disease (CKD). The three anti-rFeCysC mAbs (3-9G, 7-7C, and 9-12F) were able to recognise native FeCysC.
