Artificial DNA sequence with optimized leader function in 5′ (5′-UTR) for the improved expression of heterologous proteins in plants
A nucleotide leader sequence 5′-UTR comprises elements favorable to gene expression, such as repeated CAA trinucleotide elements in combination with repeated CT dinucleotide elements.
1. A 5′-UTR leader sequence comprising SEQ ID NO: 1.
2. A DNA construct comprising the 5′-UTR leader sequence of claim 1 operably linked upstream to a nucleic sequence coding for a heterologous protein, wherein said 5′-UTR Leader sequence is effective for enhancing expression of said heterologous protein in plants.
3. A plant expression vector comprising a 5′-UTR leader region, wherein said 5′-UTR leader region comprises the sequence shown in SEQ ID NO: 1.
4. The sequence of claim 1 , obtained from artificial synthesis.
5. A bacterial strain carrying plasmids containing the sequence as in claim 1 .
6. An engineered virus strain containing the sequence as in claim 1 , irrespective of the host organism.
7. A plant cell transformed with expression vectors containing the sequence as in claim 1 , under the control of a promoter selected from a group consisting of: a constitutive promoter, a tissue-specific promoter, an inducible promoter, a promoter with phase-dependent transcriptional activity, a promoter active in the chloroplast, and a promoter active in the mitochondrion.
8. A plant characterized by the transient expression of any protein in which the RNA messenger contains the sequence as in claim 1 , transient expression being taken to mean the production of said protein by means of viral vectors, agroinfiltration, particle delivery, or electroporation.
9. A plant, or progeny thereof, stably transformed with expression vectors containing the sequence as in claim 1 .
10. A method comprising expressing the DNA construct of claim 2 to synthesize said heterologous protein.
11. The method of claim 10 , wherein the heterologous protein
induces resistance to viral pathogens, bacterial pathogens, fungal pathogens, or herbicides.
12. The method of claim 10 , wherein the heterologous protein is selected from the group consisting of proteins involved in the production of secondary metabolites, proteins usable directly or indirectly as factors to identify and/or select transformed cells, and pharmaceutical proteins.
13. A method comprising expressing the DNA construct of claim 2 to synthesize at least one member selected from the group consisting of industrial enzymes, commercial proteins, orally administered vaccines, or injectable vaccines.
14. A method comprising expressing the DNA construct of claim 2 to synthesize patient-specific injectable vaccines to treat tumors of the lymphatic system.
15. The bacterial strain of claim 5 , wherein said bacterial strain is a strain of the species Escherichia coli, Agrobacterium tumefaciens or Agrobacterium rhizogenes.