Methods for coexpression of more than one gene using at...

C - Chemistry – Metallurgy – 12 – N

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C12N 15/83 (2006.01) A01H 5/00 (2006.01) C12N 15/82 (2006.01)

Patent

CA 2291803

A primary object of this invention is to provide a method which will enable to express simultaneously desired genes in vitro and in planta. This object is to be accomplished by utilising crTMV RNA sequences upstream of MP gene is termed here as IRESmp. The method of this invention involves the construction of recombinant DNA molecule which comprises of a transcriptional promoter, the first plant-expressible gene linked to the said transcriptional promoter, IRESmp located 3' to the first gene and the second plant-expressible gene located 3' to IRESmp such that the second gene is placed under the translational control of IRESmp. The primary chimeric continuous RNA transcript in positive sense polarity is produced by the transformed cells from plant-expressible promoter. The expression of the first gene takes place by direct translation of the 5'-proximal gene of this mRNA but the translation of the 5'-distal gene of dicistronic mRNA will be promoted by IRESmp.

L'invention concerne un procédé permettant d'exprimer simultanément les gènes désirés in vitro et in planta. Ce procédé est réalisé à l'aide de séquences d'ARN crTMV se trouvant en amont du gène MP et appelées IRESmp. Ce procédé comprend la construction d'une molécule d'ADN recombiné qui comprend un promoteur transcriptionnel, le premier gène pouvant être exprimé dans les plantes lié au promoteur transcriptionnel, IRESmp situé à la position 3' par rapport au premier gène, et le second gène pouvant être exprimé dans les plantes étant situé à la position 3' par rapport à IRESmp de sorte que le second gène soit placé sous la commande translationnelle d'IRESmp. Le produit de transcription d'ARN continu, chimère, primaire, en polarité sens positif est obtenu par les cellules transformées provenant du promoteur pouvant être exprimé dans les plantes. L'expression du premier gène s'effectue par translation directe du gène 5'-proximal de cet ARNm, mais la translation du gène 5'-distal de l'ARNm dicistronique sera activée par IRESmp.

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