Construction of recombinant adenovirus vector with SCL gene by AdMax system

【Objective】 Human SCL gene were constructed into recombinant adenovirus expression vector by AdMax vector system in order to research SCL gene function in ICC-like cells.【Methods】 SCL gene was amplified from plasmid with SCL gene by PCR and inserted to the polyclonal site of adenovirus shuttle plasmid pDC315-EGFP.HEK 293 cells were co-transfected with the constructed recombinant shuttle plasmid pDC315-EGFP/SCL and large adenovirus helper plasmid pBHGlox(delta) E1,3Cre in mediation of liposome.The obtained replication-defective recombinant adenovirus pDC315-SCL was propagated in HEK 293 cells,purified pDC315-SCL plasmid and determined for virus titer.【Results】 PCR analysis and Western Blot inspection confirmed that the human SCL gene was successfully inserted into the adenovirus vector.The titer of the recombinant adenovirus was 1×1010 PFU/mL.【Conclusions】 Recombinant adenovirus containing human SCL gene was successfully constructed by AdMax vector system.