Objective: To construct a recombinant replication-deficient adenovirus for the angiogenic inhibitor, vasostatin and assay its expression in vitro.
Methods: The cDNA for vasostatin was got by PCR amplification, then it was cloned into T vector and confirmed by enzymatic analysis and direct sequencing. Subsequently the cDNA was subcloned into the shuttle plasmid, and co-transformed into 293 cells with backbone plasmid. The resulting recombinant andenovirus was confirmed by PCR and western blot. The activity was assayed by inhibition of HUVEC growth.
Results: The PCR product was about 590 bp in length, and sequencing result was identical to that reported. The construction of recombinant replication-deficient adenovirus was confirmed by PCR. Western blot analysis showed the expression of vasostatin in vitro. The supernatant from transduced HeLa cells inhibited the growth of HUVEC specifically.
Conclusion: The recombinant adenovirus for vasostatin efficiently mediated the expression of the protein in vitro.