Construction of Recombinant Adenovirus Vector Expressing the Mouse Endostatin

Objective To construct the recombinant adenovirus vector expressing the mouse endo-statin and provide the basis for following experiments in vivo and in vitro. Methods The mEndostatin DNA which was extracted from vector containing mouse endostatin gene was successfully amplified by using PCR and then cloned into adenovirus vector pCA13. Recombinant adenovirus plasmid pCA13-mEndo was co-transfected with pBHGS into 293 packaging cells by lipofectamine2000 and the replication-deficient recombinant adenovirus Ad-mEndo was generated efficiently by homologous recombination. The Ad-mEndo recombinant adenovirus was efficiently duplicated in 293 cells and was purified by CsCl density centrifugation and the titer was measured. Results The mEndostatin cDNA was confirmed by sequencing. The virus titer was 6.3×1010pfu/ml. Conclusion This investigation provides the basis for studying tumor gene therapy by endostatin.