Construction of eukaryotic expression vector carrying HsPDF gene and its expression in NIH3T3 cells

Aim:To construct an eukaryotic expressing vector carrying Homo sapiens peptide deformylase (Hs-PDF) gene and to investigate the expression and the function of HsPDF in NIH3T3 cells.Methods:The HsPDF gene was cloned from HeLa cells by RT-PCR and then inserted into pcDNA3.1(-) eukaryotic expressing vector.The recombinant plasmid was transfected into NIH3T3 cells using positive ion liposome method before stable cell lines were established by screening with G418.The transcription of HsPDF in the transfected cells was examined by RT-PCR.MTT assay was used to evaluate the effects of HsPDF on the proliferation.Cytomorphological analysis was performed by hematoxylin-easin (HE) staining.Results:The HsPDF gene containing eukaryotic expressing vector was successfully constructed.The transcription and expression of HsPDF in the transfected cells were confirmed.High growth rate of NIH3T3 cells transfected with pcDANA3.1(-)-HsPDF was shown and several phenotypes of transformed cells were obtained.Conclusion:The HsPDF gene with its functions was expressed in NIH3T3 cells.