Methods for coexpression of more than one gene using at least one internal ribosome entry site (IRES)

• Main Authors: Atabekov, Joseph, Korpela, Timo, Dorokhov, Yurii, Ivanov, Peter, Skulachev, Maxim, Rodionova, Nina, Karpova, Olga
• Format: Patent
• Language: English
• Published: 23.04.2002

This invention relates to plant molecular biology in general, and in particular, to nucleic acid sequences which regulate the internal and 3′-proximal gene expression in polycistronic mRNA transcripts. This invention will enable the control of transgene expression through the generation of polycistronic fusion mRNAs in which all the genes are translationally active due to the presence of the IRESmp element(s). The present invention is drawn to a recombinant nucleic acid having (a) a transcriptional promoter; (b) a first plant-expressible structural gene linked to the transcriptional promoter, (c) a cDNA sequence element having an internal ribosome entry site of tobamovirus moving protein (MP) gene (IRES) which is located 3′ to the first plant-expressible structural gene, and (d) a second plant-expressible gene located 3′ to the (IRES) such that the second gene is placed under the translational control of (IRES); wherein the first plant-expressible gene, (IRES) and the second plant-expressible gene are transcribed under the action of the transcriptional promoter to give a primary transcript, and the first plant expressible gene of the primary transcript is able to translate by ribosome scanning mechanism and the second plant expressible gene of the primary transcript is capable of translation under the action of (IRES). The present invention is further drawn to a method of simultaneously expressing desired genes in vitro and in planta by using crTMV RNA sequences upstream or the MP gene, i.e. IRES.