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Biotecnologia Aplicada
Elfos Scientiae
ISSN: 0684-4551
Vol. 13, No. 1, 1996
Bioline Code: ba96005
Full paper language: English
Document type: Research Article
Document available free of charge

Biotecnologia Aplicada, Vol. 13, No. 1, 1996

 es
Vazquez, Raysa; Brito, Jose & Guerra, Maribel

Resumen

La enzima de restriccion Sac I se aislo del microorganismo Streptomyces achromogenes y se purifico a homogenidad, libre de nucleasas contaminantes. Empleando la cromatografia de intercambio ionico (Q-sefarosa y fosfocelulosa P-11), la Sac I pudo ser purificada con alto grado de pureza y usada en experimentos de clonaje. La utilidad practica de esta enzima es su gran estabilidad, alto rendimiento, facil manipulacion de las celulas productoras y la habilidad de reconocer secuencias, tales como GAGCTC. El peso molecular de esta enzima se estimo por cromatografia liquida de alta resolucion y por electroforesis en geles de acrilamida, siendo de alrededor de 50 kDa. La preparacion enzimatica fue estable al menos 20 meses, según el ensayo de estabilidad acelerado.

Palabras-clave
Endonucleasas de restriccion, purificacion, Sac I

 
 en Purification of the restriction endonuclease sac I, free of sac II and sac III contaminants
Vazquez, Raysa; Brito, Jose & Guerra, Maribel

Abstract

The restriction endonuclease Sac I was isolated from Streptomyces achromogenes and was purified to homogeneity, until no contaminant nuclease activities were detected. On the basis of ion exchange chromatography (Q-Sepharose and phosphocellulose P-11), Sac I can be obtained with a high level of purity and used in molecular cloning. The practical utitlity of Sac I enzyme is given by a high stability, high yield, easy handling of producing cells, and the ability to recognize new sequences, such as GAGCTC. The molecular weight (MW) of this enzyme was estimated by High Performance Liquid Chromatography and SDS polyacrylamide gel electrophoresis, being of about 50 kDa approximately. According to the results obtained from the accelerated stability study, the enzyme preparation is stable for at least 20 months.

Keywords
Restriction endonucleases, purification, Sac I

 
© Copyright 1996 Elfos Scientiae
Alternative site location: http://elfosscientiae.cigb.edu.cu/Archivo.asp?Id=6

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