Other

Atabekov, Joseph

Karpova, Olga

Ivanov, Peter

Korpela, Timo

Dorokhov, Yurii

Rodionova, Nina

Skulachev, Maxim

Olga Karpova

Timo Korpela

Maxim Vladimirovich Skulachev

Joseph Atabekov

Yurii Dorokhov

Peter Ivanov

Nina Rodionova

Maxim Skulachev

A primary object of this invention is to provide a method which will enable to coexpress simultaneously two (or more) desired genes in plant, animal or yeast cells, in transgenic plants and animals, or in vitro, in plant cell-derived or animal cell-derived translation systems. This objection is to be accomplished by utilizing sequence elements derived from RNAs of a tobamovirus upstream of MP gene or CP gene termed here as IRESand IRES, respectively. The method of this invention involves the construction of a recombinant nucleic acid sequence which comprises a specific transcriptional promoter, a first gene expressible in eukaryotic cells linked to said transcriptional promoter, IRESor IRESlocated 3' to the first gene and a second gene expressible in eukaryotic cells, located 3&#x2032; to IRES sequence such that the second gene is placed under the transcriptional control of IRES sequence originated from tobamovirus genome. The primary chimeric RNA transcript in positive sense polarity is produced by the transformed cells from the said promoter. The expression of the first gene occurs by direct translation whereas the translation of the 5&#x2032;-distal gene(s) of bicistronic (or polycistronic) mRNA will be promoted by IRESor IRES.

Methods for coexpression of more than one gene in eukaryotic cells

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&prime; to the first plant-expressible structural gene, and (d) a second plant-expressible gene located 3&prime; 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 .

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

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Moscow, Russia

Olga Karpova