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Objective To construct a hTERT and Cox-2 promoter regulated oncolytic virus by replacing the endogenous adenovirus E1a and E1b promoter, respectively. Methods The promoter of hTERT and Cox-2 was subcloned from human genomic DNA, then ligated to shuttle vector pd306 which contains the E1a and E1b gene of adenovirus to control the expression of E1a and E1b, respectively. The constructed shuttle vector was cotransfected with adenovirus bone vector in Ad293 cell to gain oncolytic adenovirus and determine its titer. Oncolytic virus was used to infect Hela cells to detect its execution to tumor cells. Result (1) The promoter of hTERT and Cox-2 was subcloned. (2) The hTERT and Cox-2 promoter controlled oncolytic adenovirus was gained. (3)The gained virus had ability to kill Hela cells. Contrutions The hTERT and Cox-2 dependent oncolytic adenoirus has been constructed for further research of effection of oncolytic virus on tumor cells and normal cells.
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Objective To construct a hTERT and Cox-2 promoter regulated oncolytic virus by replacing the endogenous adenovirus E1a and E1b promoter, respectively. Methods The promoter of hTERT and Cox-2 was subcloned from human genomic DNA, then ligated to shuttle vector pd306 which contains the E1a and E1b gene of adenovirus to control the expression of E1a and E1b, respectively. The constructed shuttle vector was cotransfected with adenovirus bone vector in Ad293 cell to gain oncolytic adenovirus and determine its titer. Oncolytic virus was used to infect Hela cells to detect its execution to tumor cells. Result (1) The promoter of hTERT and Cox-2 was subcloned. (2) The hTERT and Cox-2 promoter controlled oncolytic adenovirus was gained. (3)The gained virus had ability to kill Hela cells. Contrutions The hTERT and Cox-2 dependent oncolytic adenoirus has been constructed for further research of effection of oncolytic virus on tumor cells and normal cells.
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