Protein expression of human neuron-specific enolase and its antiserum preparation / 南方医科大学学报
Journal of Southern Medical University
;
(12): 1543-1547, 2006.
Artigo
em Inglês
| WPRIM
| ID: wpr-232842
ABSTRACT
<p><b>OBJECTIVE</b>To clone and express human neuron-specific enolase (HuNSE) protein and prepare NSE-specific antibody for prion disease diagnosis.</p><p><b>METHODS</b>HuNSE gene was amplified by RT-PCR and subcloned into a HIS-tagged expression vector pQE30 after sequence verification. HIS-NSE fusion protein expression was obtained in E. coli M15 after IPTG induction followed by purification of the fusion protein by Ni-NTA affinity chromatography. Two male rabbits were immunized for 4 times with the purified protein, and the antiserum against NSE protein was collected and evaluated by enzyme-linked immunosorbent assay (ELISA), Western blotting and immunohistochemistry.</p><p><b>RESULTS</b>SDS-PAGE assay yielded an approximately 22 kD HIS-NSE fusion protein. The prepared antiserum could recognize both recombinant NSE protein and native NSE protein extracted from the brain tissues of different mammalian species as shown by Western blotting and immunohistochemistry.</p><p><b>CONCLUSION</b>High expression of HuNSE is obtained in E. coli and the prepared antiserum against HuNSE can be used potentially for diagnosis of prion-associated diseases and other nervous degeneration diseases.</p>
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Fosfopiruvato Hidratase
/
Sangue
/
Proteínas Recombinantes de Fusão
/
Ensaio de Imunoadsorção Enzimática
/
Imuno-Histoquímica
/
Western Blotting
/
Clonagem Molecular
/
Doenças Priônicas
/
Diagnóstico
/
Alergia e Imunologia
Tipo de estudo:
Estudo diagnóstico
Limite:
Animais
/
Humanos
/
Masculino
Idioma:
Inglês
Revista:
Journal of Southern Medical University
Ano de publicação:
2006
Tipo de documento:
Artigo
Similares
MEDLINE
...
LILACS
LIS