AIM:To construct adenoviral vectors for luciferase driven by human telomerase reverse transcriptase(hTERT) core promoter with multimer of MYC responsive elements.METHODS:Multimer of MYC responsive elements was cloned ...AIM:To construct adenoviral vectors for luciferase driven by human telomerase reverse transcriptase(hTERT) core promoter with multimer of MYC responsive elements.METHODS:Multimer of MYC responsive elements was cloned into the upstream site of hTERT core promoter and the modified hTERT promoter and luciferase were cloned into the plasmid pDC316 to construct shuttle plasmids which cotransfected HEK 293 cells with rescue plasmid pBHGlox(delta)E1,3Cre to achieve recombinant adenoviral vectors.The cytopathic effects and PCR using primers specific for luciferase were used to identify the recombinant adenoviral vectors.RESULTS:Adenoviral vectors with luciferase driven by hTERT core promoter with none or positive and negative six copies of MYC responsive elements were constructed and amplified.The titer of the adenovirus were 3.5×106 pfu/ml,2.5×106 pfu/ml and 1.5×106 pfu/ml respectively determined by plaque assay.CONCLUSION:The further research on transciriptional targeting in osteosarcoma gene therapy can be done using adenoviral vectors with luciferase driven by hTERT promoter with MYC responsive elements.展开更多
文摘AIM:To construct adenoviral vectors for luciferase driven by human telomerase reverse transcriptase(hTERT) core promoter with multimer of MYC responsive elements.METHODS:Multimer of MYC responsive elements was cloned into the upstream site of hTERT core promoter and the modified hTERT promoter and luciferase were cloned into the plasmid pDC316 to construct shuttle plasmids which cotransfected HEK 293 cells with rescue plasmid pBHGlox(delta)E1,3Cre to achieve recombinant adenoviral vectors.The cytopathic effects and PCR using primers specific for luciferase were used to identify the recombinant adenoviral vectors.RESULTS:Adenoviral vectors with luciferase driven by hTERT core promoter with none or positive and negative six copies of MYC responsive elements were constructed and amplified.The titer of the adenovirus were 3.5×106 pfu/ml,2.5×106 pfu/ml and 1.5×106 pfu/ml respectively determined by plaque assay.CONCLUSION:The further research on transciriptional targeting in osteosarcoma gene therapy can be done using adenoviral vectors with luciferase driven by hTERT promoter with MYC responsive elements.
