2009•Journal of Zhengzhou UniversityRequires access
Construction of a luciferase reporter vector with resistance of neomycin
Yunyun Ma
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Abstract
Aim:To construct a luciferase reporter vector with neomycin resistance.Methods:To design and amplify neomycin resistance gene, then clone the gene into luciferase reporter vector pGL3,and obtain the luciferase reporter vector pGL3-neo.To compare the luciferase activity of cell lines transiently transfected by pGL3 and pGL3-CMV promoter, and cell line lasting transfected by pGL3-neo and pGL3-neo-CMV promoter after screening.Results: The neo gene was obtained by PCR; screening by restriction enzyme and sequencing analysis showed that the luciferase reporter vector with neomycin resistance had been constructed successfully. The cell lines lasting transfected by pGL3-neo and pGL3-neo-CMV promoter were separately obtained by G418 screening. The luciferase activity of the groups transiently transfected by pGL3 and pGL3-CMV promoter was 1 047±86 and 1 504±107,lower than that of groups transfected by pGL3-neo and pGL3-neo-CMV promoter (34 819±1 479 and 456 109±15 791)(P0.05).Conclusion:The luciferase reporter vector with resistance of neomycin has been successfully constructed,which remarkably elevates the sensibility and dependability of lucisferase reporter gene detection.