Expression and identification of type 1 diabetes associated autoantigen IA-2 / 中华医学杂志(英文版)
Chinese Medical Journal
;
(24): 524-528, 2003.
Artigo
em Inglês
| WPRIM
| ID: wpr-324398
ABSTRACT
<p><b>OBJECTIVES</b>To obtain prokaryotic expressed IA-2 recombinant protein and to identify its immunological activity.</p><p><b>METHODS</b>The complimentary DNA (cDNA) coding for the intracytoplasmic part of IA-2 (IA-2ic) was amplified from human fetal brain RNA, and was subcloned into the PinPoint Xa-1 T vector to construct recombinant expression plasmid, and was then expressed in E. coli JM109 cells as a fusion protein with a biotinylated peptide sequence at the aminoterminus. The biotinylated fusion protein was then purified by affinity chromatography and was subsequently dialyzed. Finally, its immunogenicity was evaluated by enzyme linked immunosorbent assay (ELISA).</p><p><b>RESULTS</b>The purified IA-2ic fusion protein resolved on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) as a single Coomassie brilliant blue stained band with a molecular weight of 59 kDa and its immunogenicity was confirmed by ELISA.</p><p><b>CONCLUSIONS</b>E. coli expressed IA-2ic fusion protein has immunological activity. It can be used for detection of IA-2 autoantibodies (IA-2A) and for further studies on type 1 diabetes in future.</p>
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Plasmídeos
/
Autoantígenos
/
Proteínas Recombinantes de Fusão
/
Proteínas Tirosina Fosfatases
/
DNA Complementar
/
Diabetes Mellitus Tipo 1
/
Alergia e Imunologia
/
Escherichia coli
/
Proteína Tirosina Fosfatase não Receptora Tipo 1
/
Proteínas Tirosina Fosfatases Classe 8 Semelhantes a Receptores
Tipo de estudo:
Estudo diagnóstico
Limite:
Animais
/
Humanos
Idioma:
Inglês
Revista:
Chinese Medical Journal
Ano de publicação:
2003
Tipo de documento:
Artigo
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