Construction and expression of pGEX-e23 (scFv) PE40 in Esche richia coli
- Author:
- No author available
- Journal Title:
- JOURNAL OF THE FORTH MILITARY MEDICAL UNIVERSITY
- Issue:
- 7
- DOI:
- 10.3321/j.issn:1000-2790.2001.07.002
- Key Word:
- No keyword available
Abstract： AIM To express a immunotoxin consisting of a single c hain variable fragment (scFv) specific for human oncogene c-erbB-2 product, p185 and P seudomonas exotoxin (PE40), named e23 (scFv) PE40, in Escherichia coli which could be u sed as an adjuvant therapy agent in breast cancer, gastric cancer and other malignant tumo r overexpressing c-erbB-2. METHODS e23 (scFv) PE40 fusion g ene for prokaryotic expression was constructed and inserted into a prokaryotic fusion protein expres sion vector pGEX-4T and expressed in Escherichia coli DH5α. RESULTS The DNA sequence of e23 (scfv) PE40 fusion gene was correct. A new anticipated Mr 90 000-protein band appeared on SDS-PAGE gel induced by IPTG and amounted to 15% of the total bacterial protein . CONCLUSION Fusion gene of e23 (scFv) PE40 has been successfully constructed and expressed in Escherichia coli DH5α.
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