Construction and identification of recombinant adenoviral shuttle vector with Egr-1 promoter and Endostatin gene

Objective To construct a recombinant adenoviral shuttle vector pshuttle-Egr1-Endostatin which contains endostatin and Egr-1 which acts as the radiation-sensitive promoter and can induce its downstream gene expression.Methods The endostatin gene fragment was amplificated from the fetal liver by RT-PCR.The Egr-1 gene was acquired from plasmid T-Egr1.Then the two genes were ligated to pMD19T and sequenced them.Finally,using the gene recombinant technique,the recombinant plasmid pshuttle-Egr1-Endostatin with radiation-inducible promoter Egr-1 was constructed.Results The sequence of both acquired endostatin and Egr-1 genes were in concordance with published that on Genebank confirmed by sequencing,indicating that the two genes were cloned successfully.Moreover,the recombinant plasmid pshuttle-Egr1-Endostatin was constructed correctly by PCR and restrictive digestion identification.Conclusion The endostatin gene and the Egr-1 promoter were cloned and the recombinant plasmid pshuttle-Egr1-Endostatin with radiation-sensitive promoter was constructed successfully in our research.