Expression, purification and identification of Abeta fusion protein with enhanced solubility / 南方医科大学学报
Journal of Southern Medical University
; (12): 447-450, 2010.
Article
de Zh
| WPRIM
| ID: wpr-355104
Bibliothèque responsable:
WPRO
ABSTRACT
<p><b>OBJECTIVE</b>To economically obtain the Abeta peptide for Alzheimer's disease (AD) research by expressing the Abeta peptide fused with the maltose binding protein (MBP) possessing high solubility in E.coli.</p><p><b>METHODS</b>The cDNA-coding sequence of Abeta peptide was modified by the addition of a BamH I site at the 5' end and a Hind III site at the 3' end using PCR. The modified sequence was ligated into the maltose-binding protein (MBP) fusion expression vector pMAL-c2 containing an thrombin cleavage site, which was transformed into competent E.coli DH5alpha cells. After identification of the single clones by PCR and DNA sequencing, the recombinant plasmid was transformed into E.coli TB1 and induced to express MBP-Abeta fusion protein. The expressed fusion protein was purified using amylose resin column and identified by SDS-PAGE and Western blotting.</p><p><b>RESULTS</b>The result of DNA sequencing verified the consistency between the inserted sequence and Abeta (1-42) sequence. SDS-PAGE electrophoresis showed that MBP-Abeta fusion protein was highly expressed in E.coli TB1, and Western blotting demonstrated that the purified fusion protein and the separated Abeta peptide could be recognized by specific anti-Abeta (22-35) antibody.</p><p><b>CONCLUSION</b>MBP-Abeta fusion protein highly expressed in E. coli TB1 cells with enhanced solubility and the separated Abeta peptide with good immunogenicity obtained may lend support to AD research.</p>
Texte intégral:
1
Indice:
WPRIM
Sujet Principal:
Protéines de fusion recombinantes
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Peptides bêta-amyloïdes
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Escherichia coli
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Protéines de liaison au maltose
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Maladie d'Alzheimer
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Vecteurs génétiques
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Génétique
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Métabolisme
Type d'étude:
Diagnostic_studies
Limites du sujet:
Humans
langue:
Zh
Texte intégral:
Journal of Southern Medical University
Année:
2010
Type:
Article