[Construction of a multi-copy Pichia expression vector and expression of human brain-derived neurotrophic factor in Pichia pastoris].
Wei Sheng Wu Xue Bao
; 45(1): 34-8, 2005 Feb.
Article
em Zh
| MEDLINE
| ID: mdl-15847159
ABSTRACT
A pPICZ alpha1 vector was reconstructed with pPICZ alpha vector and PCR product of His4-Kan sequence from pPIC9K vector. The sequence encoding human brain-derived neurotrophic factor (hBDNF) was inserted into pPICZ alpha1. The expression vectors carrying 1, 2, 3, 6 copies of expression cassettes were constructed and transformed into Pichia pastoris GS115 respectively. The transformants were screened with G418/Zeocin and induced by 0.5% methanol, and the results of SDS-PAGE confirmed that the expression protein was about 14 kD and the percentage of the protein was about 20% of the total secreted protein. ELISA and Western blot analysis demonstrated that the protein could be specifically combined with chicken antibodies to human-BDNF.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Pichia
/
Proteínas Recombinantes
/
Fator Neurotrófico Derivado do Encéfalo
/
Vetores Genéticos
Limite:
Humans
Idioma:
Zh
Revista:
Wei Sheng Wu Xue Bao
Ano de publicação:
2005
Tipo de documento:
Article
País de afiliação:
China