Recombinant human brain myelin basic protein and its antibody / 生物医学工程学杂志
Journal of Biomedical Engineering
;
(6): 64-67, 2003.
Artigo
em Chinês
| WPRIM
| ID: wpr-311107
ABSTRACT
We constructed the expression vector by inserting 21.5 KDa MBP human brain full-length cDNA coding sequence digested with restriction enzyme EcoR I and Sal I into downstream of pGEX-5T expression vector. The recombinant vector p5TMP was transformed into E. coli and the positive clonies were selected and incubated in LB medium induced by IPTG (isopropyl- -D-thiogalactoside). A new polypeptide band with apparent molecular weight 42 KDa was detected in transformed cell lysates by SDS-PAGE. Western blotting analysis confirmed that this fusion protein reacted specifically with antibodies to MBP, the expression level of MBP was about 414.6 mg/L medium estimated by immuno-dot blot, ELISA and absorbance scanning. Newzealand rabbits were immunized by subcutaneous injection of the purified recombinant MBP. The titer was obtained at 116 after 5 injections. The specificity of the antibody to MBP was confirmed by immuno-blot and Western blotting.
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Autoanticorpos
/
Proteínas Recombinantes de Fusão
/
Encéfalo
/
Expressão Gênica
/
Clonagem Molecular
/
DNA Complementar
/
Proteína Básica da Mielina
/
Alergia e Imunologia
/
Escherichia coli
/
Genética
Limite:
Animais
/
Humanos
Idioma:
Chinês
Revista:
Journal of Biomedical Engineering
Ano de publicação:
2003
Tipo de documento:
Artigo
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