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Biotecnologia Aplicada
Elfos Scientiae
ISSN: 0684-4551
Vol. 15, No. 4, 1998
Bioline Code: ba98042
Full paper language: English
Document type: Research Article
Document available free of charge

Biotecnologia Aplicada, Vol. 15, No. 4, 1998

 en Influence of the length of the P64k protein N-terminus stabilizing peptide on the expression in Escherichia coli of the fused TbpB from Neisseria meningitidis

Abstract

Biotecnología Aplicada 1998;15:254-257

Influence of the length of the P64k protein N-terminus stabilizing peptide on the expression in Escherichia coli of the fused TbpB from Neisseria meningitidis

Tamara Menéndez, Luis M Alonso, Mairet Pérez, Rolando Pajón, Ricardo Silva

Code Number: BA98042
Size of Files:
Text: 25K
Graphics: Line drawings (gif) - 6K
Photographs (jpg) - 142K

ABSTRACT

In order to improve the previously patented system for the expression of heterologous antigens in Escherichia coli, the possibility of reducing the length of the 45 amino acid (aa) stabilizer peptide from the P64k protein of Neisseria meningitidis, was investigated. The TbpB protein from N. meningitidis was expressed fused to the first 23 and 11 aa from the P64k protein and without the stabilizer peptide. The levels of expression of the TbpB protein were very low, only detectable by Western blotting when expressed without the stabilizer peptide or fused to the first 11 aa from the P64k protein. High levels of expression were achieved when the protein was expressed fused to the first 23 aa from the P64k protein. These results showed that it is possible to reduce the length of the stabilizer peptide derived from the P64k protein to one half without affecting the levels of expression of the fusion protein.

Keywords: P64k protein, transferrin binding proteins, TbpB, expression system, fusion proteins

RESUMEN

Con el objetivo de optimizar el sistema de expresión de antígenos heterólogos en Escherichia coli, patentado previamente, se investigó la posibilidad de reducir la talla del fragmento estabilizador de 45 aminoácidos (aa) provenientes de la proteína P64k de Neisseria meningitidis. La proteína TbpB de N. meningitidis se expresó en E.coli fusionada a los primeros 23 y 11 aa de la proteína P64k y sin el fragmento estabilizador. En los dos últimos casos se obtuvieron niveles bajos de expresión de TbpB, solo detectables mediante Western blotting. Se obtuvieron altos niveles de expresión de TbpB, cuando se expresó fusionada a los primeros 23 aa de la proteína P64k. Estos resultados muestran que es posible reducir hasta la mitad la talla del fragmento estabilizador derivado de la proteína P64k sin afectar los niveles de expresión de la proteína de fusión.

Palabras claves: proteína P64k, proteínas de unión a transferrina, TbpB, sistemas de expresión, proteínas de fusión

Copyright 1998 Elfos Scientiae

 

Alternative site location: http://elfosscientiae.cigb.edu.cu/Archivo.asp?Id=6

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