Por favor, use este identificador para citar o enlazar este ítem: http://repositorio.ufc.br/handle/riufc/59741
Tipo: Artigo de Periódico
Título : Cloning of serine protease cDNAs from Crotalus durissus terrificus venom gland and expression of a functional Gyroxin homologue in COS7 cells
Título en inglés: Cloning of serine protease cDNAs from Crotalus durissus terrificus venom gland and expression of a functional Gyroxin homologue in COS7 cells
Autor : Yonamine, C.M.
Prieto-da-Silva, A.R.B.
Magalhães, G. S.
Rádis-Baptista, Gandhi
Morganti, Ligia
Ambiel, F. C.
Chura-Chambi, R.M.
Camillo, M.A.P.
Palabras clave : Protease;Enzimas
Fecha de publicación : 2009
Editorial : Toxicon.
Citación : YONAMINE, C.M.; PRIETO-DA-SILVA, A.R.B.; MAGALHÃES, G.S.; RÁDIS-BAPTISTA, Ghandi; Morgant,L.; AMBIEL, F.C.; Chura-Chambia, R.M.; YAMANE, T.; CAMILLO, M.A.P. Cloning of serine protease cDNAs from Crotalus durissus terrificus venom gland and expression of a functional Gyroxin homologue in COS7 cells. Toxicon, United Kingdom, v.54, p. 110–120. 2009.
Abstract: Gyroxin is one of main serine proteases of Crotalus durissus terrificus venom, representing about 2% of the protein content in the crude venom. It is a 33 kDa glycoprotein with 3.8% by weight of sugar moiety. This toxin induces hemotoxicity in mice and a neurological condition called barrel rotation syndrome. In the present work, we report the molecular cloning of five new nucleotide sequences from a cDNA library of the venom glands of a single specimen of C. d. terrificus. These sequences have been analyzed in silico with respect to their cDNA organization and similarity with other snake venom serine proteases (SVSPs). We also describe a rapid and efficient method for screening vectors for mammalian cell expression, based on the fact that SVSPs are difficult-to-express toxins due to the presence of several disulfide bonds and glycosylation in their structures. Thus, one of the Gyroxin cDNAs was subcloned into pSectag2 HygroA and pED vectors and used to transfect COS-7 cells. Expression of the functional recombinant Gyroxin isoform was achieved with this cell line with esterase activity in the conditioned culture medium, as revealed by immunoblot of secreted protein and standard anti-crotalic serum from Butantan Institute.
URI : http://www.repositorio.ufc.br/handle/riufc/59741
ISMN : 0041-0101
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