Construction and Characterization of EBNA3C and RASSF1A Plasmids

The plasmids pA3F-EBNA3C, pA3F-EBNA3C 1–365, pA3F-EBNA3C 366–620 and pA3F-EBNA3C 621–992 encoding full-length Flag-tagged EBNA3C and truncated mutations of EBNA3C have been described previously [65 (link)]. The full-length RASSF1A was amplified from total RNA by reverse transcription PCR via using primers 5’-CGCGGATCCATGTC-GGGGGAGCCTGAG-3’ and 5’-CCGCTCGAGACTCCCCAGAGTCATTTTCCTTCAGG-3’. The PCR products were cloned into pA3M and pEGFP-N1, respectively. The pA3M-RASSF1A cDNA was used as the template for PCR amplification to generate RASSF1A mutants. The PCR products were cloned into pA3M. pGL4.2-R1A-promoter used to express RASSF1A promoter was amplified from genomic DNA by primers: 5’-GAAGATCTAGCCCAGGGTGACAGAGCCA-AATG-3’ and 5’-CCAAGCTTGGCCCGGTTGGGCCCGTGCTTC-3’. PCR products were inserted into pGL4.20[luc2/Puro] vector (Promega Corporation, Madison, USA) by Bgl II and Hind III.