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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.
Assuntos
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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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