Objective: To investigate the possibility of constructing eukaryotic expression vector for human angiopoietin 1 (hAng-1), transfecting it to bone marrow mesenchymal stem cells (MSCs) so as to repair bone defect.
Methods: The eukaryotic expression vector pcDNA3-hAng-1 was constructed by recombinant DNA technique, transfected into MSCs by liposome DOTAP, and selected with G418. The hAng-1 expression of mRNA and protein was detected by reverse transcript-PCR and Western Blot.
Results: After the recombinant eukaryotic expression vector for hAng-1 was digested with Xho-I and BamH-I, electrophoresis revealed 1.4 kb fragment for hAng-1 gene and 5.4 kb fragment for pcDNA3 vector. In the transfected MSCs, the mRNA and protein expression of hAng-1 gene were detected with reverse transcript-PCR and Western Blot.
Conclusion: The constructed eukaryotic expression vector hAng-1 could be expressed in the transfected MSCs, thus to provide the basis for bone repair with tissue engineering.