Aim To construct and identified the eukaryotic expression recombinant plasmids containing caveolin-1 gene and mutants for the potential functional analysis. Methods Using a novel high fidelity gene expression profiling strategy mediated by Pfu proofreading polymerases combining with phosphorothioate modified primers, PCR-amplified ORFs of caveolin-1 and its mutants were inserted into a N-GFP tagging expression vector with the help of topoisomerase I-mediated ligation. After transformation into chemically competent One Shot TOP10 E.coli,the respective destination clones were selected for by plating on ampicillin selective agar plates. The correct orientation and reading frames of the constructs were identified through the PCR approach with the respective sense primer and the BGH antisense primer. The corrected reconstructs would add about 100 bp more than the RT-PCR fragements. The integrity of ORFs was verified by sequencing to exclude errors introduced in the amplification step. MTT and western blot approaches were carried out to determine whether the overexpressed tagged proteins present the same protein activities as endogenous proteins. Results We obtained 3 recombined caveolin-1 mutant plasmids. In addition,we demonstrated that overexpression of two caveolin-1 mutants in HepG2 cells could result in apoptosis of HepG2. However the other caveolin-1 mutant could promote the growth of HepG2 cells. The overexpressed tagged proteins present the same protein activities as endogenous caveolin-1 protein. Conclusion The eukaryotic expression recombinant plasmids containing caveolin-1 gene and mutants were successfully constructed.