Comparative evaluation of heat shock and electroporation transformation of canine homlogue of HER2 gene cloned in a PUC57 plasmid construct

Document Type : Original Article

Authors

1 Department of Microbiology, Faculty of Veterinary Medicine, Cairo University.

2 Department of Biochemistry, Faculty of Veterinary Medicine, Cairo University.

Abstract

Human epidermal growth factor receptor 2 (HER2) is an important prognostic marker in the case of breast cancer and other tumors. Canines remain one of the most important models for comparative oncology with humans due to the great similarity in the spontaneous presentation and development of cancer, and in the high homology in the amino acid sequence within the HER2 gene-encoded polypeptide. In this study, the canine homolog of the HER2 DNA sequence constructed in PUC57 plasmid was transformed in DH5α E.coli by heat-shock and electroporation. The plasmid was extracted from DH5α by Miniprep protocol and the HER2 gene 3738 bp was successfully amplified from PUC57 plasmid by polymerase chain reaction (PCR). Both heat shock and electroporation were efficient and resulted in the suspected amplicon. However, electroporation gave a higher transformation rate. HER2 DNA sequence will be used for the production of the recombinant HER2 protein in the near future.

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