Key Function：co-expression of two genes of interest
Mammalian Selection：Neo，G418
Cloning Method：Restriction Enzyme/MCS

pVITRO is a family of plasmids developed mainly for in vitro studies. They allow the ubiquitous and constitutive co-expression of two genes of interest.

pVITRO plasmids can be stably transfected in mammalian cells and the genes of interest are expressed at high levels.

Each pVITRO plasmid is available with either two multiple cloning sites or two reporter genes.

Benefits

• More efficient and convenient than using two separate vectors
• High levels of expression from two separate transcription units
• A choice of selectable markers that are active both in E. coli and mammalian cells
• Convenient cloning of cDNAs with two multiple cloning sites (MCS)
• Or also available with GFP and LacZ Reporter Genes

Gene Combinations in a Single Plasmid

Co-expression of two or more genes from a single vector is more efficient and convenient than using two separate vectors. pVITRO are multigenic plasmids that contain two distinct transcription units (TU).

Strong and Constitutive Expression of Two Transgenes

Each pVITRO plasmid features two constitutive promoters that drive high levels of expression from two separate TU in a large number of mammalian cell lines.

Transcriptional interference is minimized by using promoters of different origins or promoters that are coordinately activated (i.e. ferritin promoters), and strong polyadenylation signals (polyA).

• pVITRO1 plasmids carry two elongation factor 1 alpha (EF1α) promoters, from rat and mouse origins combined to the CMV and SV40 enhancers respectively.
• pVITRO2 plasmids contain two human ferritin composite promoters, FerH (heavy chain) and FerL (light chain), combined to the SV40 and CMV enhancers respectively. To eliminate the iron regulation, their 5‘UTRs have been replaced by the 5‘UTR of the mouse and chimpanzee EF1α genes.

Single Selection Marker for E. coli and Mammalian Cells

pVITRO plasmids are available with different selectable markers that are active both in E. coli and mammalian cells: bsr (blasticidin resistance), hph (hygromycin resistance), neo (kanamycin/G418 resistance). In bacteria, the resistance gene is expressed from the E. coli EM7 promoter. In mammalian cells, it is transcribed from the promoter located 3‘ of the Ori, as a polycistronic mRNA and translated through the IRES of the Foot and Mouth Disease Virus.

Available with Two MCS or Two Reporter Genes

pVITRO-mcs plasmids contain two multiple cloning sites (MCS) for the convenient cloning of cDNAs.

pVITRO-GFP/LacZ express the GFP and LacZ reporter genes, and can be used as controls. As the reporter genes are flanked by unique restriction sites, they can be used for the cloning of open reading frames that can be chosen from InvivoGen‘s Open Reading Frame lists.

pVITRO2-mcs

pVITRO2 plasmids contain two human ferritin composite promoters, FerH (heavy chain) and FerL (light chain), combined to the SV40 and CMV enhancers respectively. To eliminate the iron regulation, their 5’UTRs have been replaced by the 5’UTR of the mouse and chimpanzee EF-1α genes.

pVITRO2-mcs is available with 2 multiple cloning sites (MCS).

• MCS 1 5‘- Age I, Eco RV, Bam HI, Mlu I, Cla I, Sal I, Avr II -3‘
• MCS 2 5‘- Sgr AI, Fsp I, Bgl II, Eco RI, Bst BI, Xho I, Nhe I-3‘