Insert sequence length determines transfection efficiency and gene expression levels in bicistronic mammalian expression vectors.

ABSTRACT: Bicistronic expression vectors have been widely used for co-expression studies since the initial discovery of the internal ribosome entry site (IRES) about 25 years ago. IRES sequences allow the 5' cap-independent initiation of translation of multiple genes on a single messenger RNA strand. Using a commercially available mammalian expression vector containing an IRES sequence with a 3' green fluorescent protein fluorescent marker, we found that sequence length of the gene of interest expressed 5' of the IRES site influences both expression of the 3' fluorescent marker and overall transfection efficiency of the vector construct. Furthermore, we generated a novel construct expressing two distinct fluorescent markers and found that high expression of one gene can lower expression of the other

SUBMITTER: Payne AJ 

PROVIDER: S-EPMC3867706 | biostudies-literature | 2013

REPOSITORIES: biostudies-literature