Expression, purification and characterization of a synthetic gene encoding human amyloid beta (Abeta1-42) in Escherichia coli.
Indian J Biochem Biophys
;
2007 Apr; 44(2): 71-5
Artigo
em Inglês
| IMSEAR
| ID: sea-26635
ABSTRACT
Alzheimer's disease (AD) is a neurodegenerative disorder characterized by a progressive loss of cognitive function. Existing evidence indicates that abnormal processing and extracellular deposition of the longer form of the amyloid peptide Abeta(1-42), a proteolytic derivative of the amyloid precursor protein (APP), is a key step in the pathogenesis of AD. Active immunization with Abeta(1-42) has been shown to decrease brain beta deposition and improve cognitive performance in mouse models of AD. In the present study, we sought to express the synthetic gene encoding AB in Escherichia coli to enable rapid production of the antigen and its purification. The synthetic gene has been constructed from six oligonucleotides by employing overlapping PCR strategy and expressed in E. coli using the T7 promoter system. The recombinant peptide has been purified to homogeneity by a single step Ni+2 affinity chromatography. Enzyme-linked immunosorbent assay (ELISA) using polyclonal anti-Abeta(1-42) sera confirms that the corresponding linear B-cell epitopic sequences are available for immunorecognition in the recombinant peptide. This methodology enables rapid, continuous production and purification in bulk amounts of human Abeta sequence by employing bacterial expression system
Texto completo:
DisponíveL
Índice:
IMSEAR (Sudeste Asiático)
Assunto principal:
Fragmentos de Peptídeos
/
Proteínas Recombinantes
/
Humanos
/
Ensaio de Imunoadsorção Enzimática
/
Dados de Sequência Molecular
/
Peptídeos beta-Amiloides
/
Cromatografia de Afinidade
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Sequência de Aminoácidos
/
Clonagem Molecular
/
Escherichia coli
Idioma:
Inglês
Revista:
Indian J Biochem Biophys
Ano de publicação:
2007
Tipo de documento:
Artigo
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