Cloning and characterization of promoter region of cell division autoantigen 1 / 吉林大学学报(医学版)

ABSTRACT

Objective To clone different lengths cell division autoantigen1(CDA1) promoters(CDA1P) and measure their activities in order to provide basis for fumctional study of CDA1P.Methods Different lengths CDA1P were amplified by PCR using mouse liver genomic as template and cloned into BglⅡ digested pGL3-basic plasmid to produce recombinant plasmid pGL3-basicmCDA1P,then they were transfected into Lewis lung cell line(LLC) and RAW 264.7 cells differently,these cells were collected after transfected for 48 h,finally luciferase activities of pGL3-basic-mCDA1P were measured.Results ①Different lengths CDA1P were cloned and characterized by restriction enzymes.②Different lengths CDA1P were ligated into pGL3basic vector,and were proved with PCR and DNA sequencing .③Activities of pGL3-basic-mCDA1A were different in the two kinds of Leuwis and mononuclear macrophage cells(RAW264.7 cells).Conclusion The different activities of CDA1P may be related with different cellular types.