Expression and identification of the extracellular domain of human epidermal growth factor receptor type III variant / 南方医科大学学报
Journal of Southern Medical University
;
(12): 151-153, 2008.
Article
Dans Chinois
| WPRIM
| ID: wpr-293430
ABSTRACT
<p><b>OBJECTIVE</b>To clone the extracellular domain (ECD) of the type III variant of human epidermal growth factor receptor (EGFRvIII) and construct the recombinant expression plasmid.</p><p><b>METHODS</b>A DNA fragment (vIII ECD) encoding the extracellular domain of human EGFRvIII was obtained by PCR, and its T-A was cloned and sequenced. The DNA fragment was then ligated into the GST fusion expression vector to construct the recombination plasmid. After identification with restriction digestion and DNA sequencing, the recombinant plasmid was transformed into E. coli BL21 (DE3) for expression of the recombinant protein. The target protein was identified by SDS-PAGE and Western blotting.</p><p><b>RESULTS</b>The results of restriction digestion and DNA sequencing confirmed the successful construction of the recombinant plasmid. SDS-PAGE showed that the fusion protein was expressed as inclusion bodies in E. coli BL21 (DE3), and the amount of the fusion protein expressed in the bacteria, after induction for 4 h, accounted for up to 15% of the total bacterial proteins. Western blotting demonstrated that the fusion protein could be recognized by the specific anti-EGFR antibody.</p><p><b>CONCLUSION</b>We have successfully constructed the recombinant expression vector of vIII ECD and induced the expression of the fusion protein, which may facilitate functional and immunological studies of EGFRvIII.</p>
Texte intégral:
Disponible
Indice:
WPRIM (Pacifique occidental)
Sujet Principal:
Plasmides
/
Protéines de fusion recombinantes
/
Clonage moléculaire
/
Analyse de séquence d'ADN
/
Escherichia coli
/
Récepteurs ErbB
/
Vecteurs génétiques
/
Génétique
/
Métabolisme
Type d'étude:
Etude diagnostique
/
Étude pronostique
Limites du sujet:
Humains
langue:
Chinois
Texte intégral:
Journal of Southern Medical University
Année:
2008
Type:
Article
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