Construction of plasmid bicistronic vectors and their application for in vitro transfection
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Internal ribosome entry site (IRES) sequences, which stand for the basic element of cap 5'-independent translation, are currently widely used to coexpress heterologous genes from one plasmid. In this study construction of four bicistronic plasmids containing IRES and application of these vectors for transfection of in vitro cultured cells were described. The obtained data show that constructed bicistronic plasmids are very efficient in vitro in terms of simultaneous expression of fibroblast growth factor-4 (FGF-4) and vascular endothelial growth factor (VEGF) or one of these factors and green fluorescent protein (GFP) from one plasmid. Interestingly, expression of two genes, although simultaneous, is not equal. It has turned out that IRES-dependent mRNA translation is less efficient than cap 5'-dependent translation of the first gene, which should be taken into account during construction of bicistronic plasmids.
Publication order reference
Jozef Dulak, Zaklad Biotechnologii Medycznej, Wydzial Biochemii, Biofizyki i Biotechnologii, Uniwersytet Jagiellonski, ul. Gronostajowa 7, 31-271 Krakow, Poland