Construction of Eukaryotic Expression Plasmid pcDNA3-FHIT and Its Transient Expression in COS-1 Cells

摘要

Objective: To construct eukaryotic expression plasmid pcDNA3-fragile histidine triad(FHIT) and obtain its transient expression in COS-1 cells. Methods: FHIT gene was cloned from normal human thyroid tissue by RT-PCR and then inserted into eukaryotic expression vector pcDNA3. After the sequence was confirmed, the recombinant plasmid pcDNA3-FHIT was transfected into COS-1 cells by cation liposome. The transient expression in the cells was measured by immunocytochemistry. Results: The sequence of FHIT in pcDNA3 was correct and high expression was obtained in COS-1 cells. Conclusion: The eukaryotic expression plasmid pcDNA3-FHIT was constructed successfully and could highly express FHIT protein in COS-1 cells. This will be potentially useful for the research on gene therapy.