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[Expression and purification of gene transfer vehicle mediated by epidermal growth factor receptor]
Hua-sheng Fang1, Lin Zhang, Sheng-dong Lu
1National Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, CAMS, PUMC, Beijing 100005, China.
Objective:
To create a gene transfer vehicle for targeting gene therapy of cancer with epidermal growth factor receptor overexpressing.
Methods:
Encoding sequences of the first domain of histone gene (H1) and EGF C-loop (EGFc) were obtained by PCR amplification. These two DNA fragments were ligated by EcoR I site, and cloned and sequenced. E. coli expression vector of the fusion gene was then constructed. The fusion protein H1EGFc was purified by specific band isolation from SDS-PAGE.
Results:
The molecular weight of purified protein was consistent with the designed request. Its purity reached 94.02%.
Conclusion:
A fusion protein H1EGFc was expressed and purified.