Figure 4From: Rapid obtention of stable, bioluminescent tumor cell lines using a tCD2-luciferase chimeric constructStructure of monocistronic and bicistronic constructs inserted in lentiviral vectors for combined expression of the luciferase 2 and truncated rat CD2 proteins. Lentiviral vectors were designed for expression of the luciferase 2 protein combined with a truncated form of the rat CD2 (tCD2) protein either as a fusion protein (monocistronic insert) or as two distinct proteins (bicistronic insert). Both types of constructs are inserted at an EcoR1 site in the pro-lentiviral plasmid under the control of the EF-1 alpha promoter and flanked by the WPRE sequence. Recombinant lentiviral particles were produced by tri-transfection in 293T cells as explained in the Materials and Methods section.A) The tCD2-luc2 insert contains the first 232 codons of the rat CD2 gene fused to 2 codons GAA (glutamic acid) and GTC (valine) added for cloning purpose (EcoR1 site) in frame with the full length luciferase 2 gene starting at codon 236.B) The bicistronic insert - luc2-IRES-tCD2 - contains the full length luciferase 2 gene linked by an IRES (Internal Ribosome Entry System sequence) to the truncated rat CD2 gene. This IRES (573 base pairs) was derived from the pQCXIX plasmid (Clontech).Back to article page