Psod expression units

Abstract

The present invention relates to the use of nucleic acid sequences for regulating the transcription and expression of genes, the novel promoters and expression units themselves, methods for altering or causing the transcription rate and/or expression rate of genes, expression cassettes comprising the expression units, genetically modified microorganisms with altered or caused transcription rate and/or expression rate, and methods for preparing biosynthetic products by cultivating the genetically modified microorganisms.

1 . An isolated nucleic acid molecule having promoter activity, selected from the group consisting of
 A) a nucleic acid molecule comprising the nucleotide sequence of SEQ ID NO:1;   B) a nucleic acid molecule comprising a nucleotide sequence of at least 90% identity to the nucleotide sequence of SEQ ID NO:1;   C) a nucleic acid molecule which hybridizes with the complement of the nucleotide sequence of SEQ ID NO:1; and   D) a nucleic acid molecule comprising a fragment of the nucleic acid molecule of (A), (B) or (C),   
       wherein the molecule has promoter activity; wherein the nucleic acid molecule does not consist of SEQ ID NO:1. 
     
     
         2 . A method of regulating the transcription of a gene comprising introducing into a host cell the nucleic acid molecule of  claim 1  or a nucleic acid molecule consisting of SEQ ID NO:1, wherein the nucleic acid molecule has promoter activity. 
     
     
         3 . A method for altering or causing the transcription rate of genes in microorganisms compared with the wild type by
 a) altering the specific promoter activity in the microorganism of endogenous nucleic acids having promoter activity, which regulate the transcription of endogenous genes, compared with the wild type or   b) regulating the transcription of genes in the microorganism by nucleic acids having promoter activity or by nucleic acids having promoter activity with altered specific promoter activity according to embodiment a), where the genes are heterologous in relation to the nucleic acids having promoter activity.   
     
     
         4 . The method according to  claim 3 , wherein the regulation of the transcription of genes in the microorganism by nucleic acids having promoter activity or by nucleic acids having promoter activity with altered specific promoter activity according to embodiment a) is achieved by
 a) introducing one or more nucleic acids having promoter activity, where appropriate with altered specific promoter activity, into the genome of the microorganism so that transcription of one or more endogenous genes takes place under the control of the introduced nucleic acid having promoter activity, where appropriate with altered specific promoter activity, or   b) introducing one or more genes into the genome of the microorganism so that transcription of one or more of the introduced genes takes place under the control of the endogenous nucleic acids having promoter activity, where appropriate with altered specific promoter activity, or   c) introducing one or more nucleic acid constructs comprising a nucleic acid having promoter activity, where appropriate with altered specific promoter activity, and functionally linked one or more nucleic acids to be transcribed, into the microorganism.   
     
     
         5 . The method according to  claim 3  or  4 , wherein to increase or cause the transcription rate of genes in microorganisms compared with the wild type
 a) the specific promoter activity in the microorganism of endogenous nucleic acids having promoter activity, or which regulate the transcription of endogenous genes, is increased compared with the wild type, or   b) the transcription of genes in the microorganism is regulated by nucleic acids having promoter activity or by nucleic acids having increased specific promoter activity according to embodiment a), where the genes are heterologous in relation to the nucleic acids having promoter activity.   
     
     
         6 . The method according to  claim 5 , wherein the regulation of the transcription of genes in the microorganism by nucleic acids having promoter activity or by nucleic acids having promoter activity with increased specific promoter activity according to embodiment a) is achieved by
 a) introducing one or more nucleic acids having promoter activity, where appropriate with increased specific promoter activity, into the genome of the microorganism so that transcription of one or more endogenous genes takes place under the control of the introduced nucleic acid having promoter activity, where appropriate with increased specific promoter activity, or   b) introducing one or more genes into the genome of the microorganism so that transcription of one or more of the introduced genes takes place under the control of the endogenous nucleic acids having promoter activity, where appropriate with increased specific promoter activity, or   c) introducing one or more nucleic acid constructs comprising a nucleic acid having promoter activity, where appropriate with increased specific promoter activity, and functionally linked one or more nucleic acids to be transcribed, into the microorganism.   
     
     
         7 . The method according to  claim 3  or  4 , wherein to reduce the transcription rate of genes in microorganisms compared with the wild type
 a) the specific promoter activity in the microorganism of endogenous nucleic acids having promoter activity, which regulate the transcription of endogenous genes, is reduced compared with the wild type, or   b) nucleic acids having reduced specific promoter activity according to embodiment a) are introduced into the genome of the microorganism so that the transcription of endogenous genes takes place under the control of the introduced nucleic acid having reduced promoter activity.   
     
     
         8 . The method according to  claim 3 , wherein the genes are selected from the group of nucleic acids encoding a protein from the biosynthetic pathway of proteinogenic and non-proteinogenic amino acids, nucleic acids encoding a protein from the biosynthetic pathway of nucleotides and nucleosides, nucleic acids encoding a protein from the biosynthetic pathway of organic acids, nucleic acids encoding a protein from the biosynthetic pathway of lipids and fatty acids, nucleic acids encoding a protein from the biosynthetic pathway of diols, nucleic acids encoding a protein from the biosynthetic pathway of carbohydrates, nucleic acids encoding a protein from the biosynthetic pathway of aromatic compounds, nucleic acids encoding a protein from the biosynthetic pathway of vitamins, nucleic acids encoding a protein from the biosynthetic pathway of cofactors and nucleic acids encoding a protein from the biosynthetic pathway of enzymes, where the genes may optionally comprise further regulatory elements. 
     
     
         9 . The method according to  claim 8 , wherein the proteins from the biosynthetic pathway of amino acids are selected from the group of aspartate kinase, aspartate-semialdehyde dehydrogenase, diaminopimelate dehydrogenase, diaminopimelate decarboxylase, dihydrodipicolinate synthetase, dihydrodipicolinate reductase, glyceraldehyde-3-phosphate dehydrogenase, 3-phosphoglycerate kinase, pyruvate carboxylase, triosephosphate isomerase, transcriptional regulator LuxR, transcriptional regulator LysR1, transcriptional regulator Lys R2, malate-quinone oxidoreductase, glucose-6-phosphate deydrogenase, 6-phosphogluconate dehydrogenase, transketolase, transaldolase, homoserine O-acetyltransferase, cystathionine gamma-synthase, cystathionine beta-lyase, serine hydroxymethyltransferase, O-acetyl homoserine sulfhydrylase, methylenetetrahydrofolate reductase, phosphoserine aminotransferase, phosphoserine phosphatase, serine acetyltransferase, homoserine dehydrogenase, homoserine kinase, threonine synthase, threonine exporter carrier, threonine dehydratase, pyruvate oxidase, lysine exporter, biotin ligase, cysteine synthase I, cysteine synthase II, coenzyme B12-dependent methionine synthase, coenzyme B12-independent methionine synthase, sulfate adenylyltransferase subunit 1 and 2, phosphoadenosine-phosphosulfate reductase, ferredoxin-sulfite reductase, ferredoxin NADP reductase, 3-phosphoglycerate dehydrogenase, RXA00655 regulator, RXN2910 regulator, arginyl-tRNA synthetase, phosphoenolpyruvate carboxylase, threonine efflux protein, serine hydroxymethyltransferase, fructose-1,6-bisphosphatase, protein of sulfate reduction RXA077, protein of sulfate reduction RXA248, protein of sulfate reduction RXA247, protein OpcA, 1-phosphofructokinase and 6-phosphofructokinase.