High expression
View Patent ↗Identified are mutants of the promoter of the Z. mobilis glyceraldehyde-3-phosphate dehydrogenase gene, which direct improved expression levels of operably linked heterologous nucleic acids. These are high expression promoters useful for expression of chimeric genes in Zymomonas, Zymobacter , and other related bacteria.
1. An isolated nucleic acid molecule comprising a Z. mobilis glyceraldehyde-3-phosphate dehydrogenase gene promoter that has a base substitution in a position selected from the group consisting of position 116, position 217, or both positions 116 and 217; wherein the position numbers are of SEQ ID NO:1; and wherein at position 116 a T replaces G, and at position 217 a T replaces C.
2. The isolated nucleic acid molecule of claim 1 comprising a sequence selected from the group consisting of SEQ ID NO:4, 5, 6, 7, 8 9, 10, 11, and 12.
3. A chimeric gene comprising the isolated nucleic acid molecule of claim 1 operably linked to a heterologous nucleic acid molecule.
4. The chimeric gene of claim 3 wherein the heterologous nucleic acid molecule encodes a protein or peptide.
5. The chimeric gene of claim 3 wherein the heterologous nucleic acid molecule codes for a regulatory RNA molecule selected from the group consisting of an antisense RNA, a ribozyme, and an interfering RNA.
6. A vector comprising the isolated nucleic acid molecule of claim 1 .
7. A vector comprising the isolated nucleic acid molecule of claim 2 .
8. A method of transforming a bacterial cell selected from the group consisting of Zymomonas cells and Zymobacter cells comprising introducing into the cell the isolated nucleic acid molecule of claim 1 .
9. A method of transforming a bacterial cell selected from the group consisting of Zymomonas cells and Zymobacter cells comprising introducing into the cell the isolated nucleic acid molecule of claim 2 .
10. The method according to claim 8 wherein introducing comprises integrating the isolated nucleic acid molecule into the genome of the cell or maintaining on a stably replicating plasmid within the cell.