EP0680511A1

EP0680511A1 - Acetyl-coa-carboxylase-gene

Info

Publication number: EP0680511A1
Application number: EP94905669A
Authority: EP
Inventors: Reinhard TÖPFER; Wolfgang Schulte; Jeff Schell
Original assignee: Max Planck Gesellschaft zur Foerderung der Wissenschaften eV
Current assignee: Max Planck Gesellschaft zur Foerderung der Wissenschaften eV
Priority date: 1993-01-22
Filing date: 1994-01-21
Publication date: 1995-11-08
Also published as: CA2150678A1; WO1994017188A3; JPH08509116A; WO1994017188A2

Abstract

A DNA-sequence which codes for the acetyl-CoA-carboxylase is disclosed, as well as the alleles and derivates of said DNA-sequence. The acetyl-CoA-carboxylase gene sequence may be used in order to obtain by heterologous expression species of plants, for example cereal plants, which are resistant to herbicides against graminaceous plants, or to modify by homologous or heterologous expression the quality and quantity of vegetable oils and fats.

Description

Acetyl-CoA carboxylase gene

The invention relates to a DNA sequence which codes for the acetyl-CoA carboxylase, and the alleles and derivatives of this DNA sequence.

The enzyme acetyl-CoA carboxylase (EC 6.4.1.2) is an important key enzyme in the fatty acid metabolism of prokaryotes and eukaryotes. In a two-step reaction, it catalyzes the ATP-dependent carboxylation of acetyl-CoA to malonyl-CoA (AW Alberts and PR Vagelos, The Enzymes (Boyer PD ed), Vol. 6, pp. 37-82, 3rd edition, Academic Press, New York, 1972) according to the following reaction equations:

BCCP + HC0 ₃ ^" biotin carboxylase BCCP-COO- + ADP + Pi

BCCP-COO ^" + acetyl-CoA transcarboxylase BCCP + malonyl-CoA Acetyl-CoA carboxylase (ACC) has been investigated biochemically, particularly in animal systems and E. coli, and molecular biological studies on a wide variety of organisms have recently been carried out, such as in the rat (F. Lopez-Casillas, DH Bai, X. Luo, IS Kong, MS Hermodson and KH Kim, PNAS 85, pp. 5784-5788 (1988)), the chicken (T. Takai, C. Yokoymama, K. Wada and T. Tanabem J. Biol Soc. 263, pp. 2651-2657, (1988)), the yeast (W. Al-Feel, SS Chirala and SJ Wakil, PNAS 89, pp. 4534-4538, (1984)) and E. coli (J -H. Alix, DNA 8: pp. 779-789 (1989); H. Kondo, K. Shiratuchi, T. Yoshimoto, T. Masuda, A. Kitazono, D. Truru, M. Anai, M. Sekiguchi and T. Tanabe, PNAS 88: pp. 9730-9733 (1991); S.-J. Li and JE Cronan, Jr., J. Biol. Chem. 267, pp. 855-863 (1992a)).

In bacteria, the ACC enzyme consists of three different polypeptide chains, which consist of three functional units consisting of the biotin carboxylase (BC), the biotin carboxy carrier protein (BCCP) and the carboxyl transferase (CT)

(H.G. Wood and R.E. Barden, Annu. Rev. Biochem. 46, pp. 385-413, (1977)). Portions of the amino acid sequence of the E. coli ACC enzyme in the region of the biotin domain were used by M.R. Sutton, R.R. Fall, A.M. Nervi, A.W. Alberts, P.R. Vagelos and R.A. Bradshaw, J. Biol. Chem. 252, pp. 3934-3940 (1977)). The genes for the BCCP and the BC from E. coli (J.-H. Alix, supra) as well as for the CT

(S.-J. Li and JE Cronan, supra) have been described recently. The molecular weights of these proteins derived from the nucleic acid sequence are 17 kD for the BCCP, 49 kD for the BC and 35 kD for the α subunit of the CT and 33 kD for the β subunit of the CT. In animals, yeast and plants, the three functional units or domains mentioned are combined in one polypeptide (DG Hardie and P. Cohen, FEBS Letters 91, pp. 1-7 (1978), M. Mishina, R. Roggenkamp and E. Schweizer, Eur. J. Biochem. 111, pp. 79-87 (1980), B. Egin-Bühler, R. Loyal and J. Ebel, Arch. Biochem. Biophys. 203, pp. 90-100, ( 1979), AR Slabas and A. Hellyer, Plant Sei 39, pp. 177-182, (1985), Hellyer et al, JL Harwood, Ann. Rev. Plant. Physiol. 39, pp. 101-138 (1988)) . The molecular weights of a multifunctional subunit are over 200 kD. The ACC of the rat has a molecular weight of 265 kD (Lopez-Casillas et al, supra), that of the yeast a molecular weight of 251 kD (Al-Feel e_t al, supra) and that of plants varies between 210 and 240 kD (Hellyer et al, supra).

Table 1 shows an overview of the homologies of the known ACC enzymes.

Table 1

Table 1 shows the percentages of identical amino acids and the degree of homology in acetyl-CoA carboxylases from chicken, from the rat, from the yeast and from E. coli. It is clearly shown that the ACC enzymes also show a relatively high degree of kinship across the various organisms. Despite the large evolutionary gap between rat and chicken on the one hand and yeast on the other hand there is still about 66% homology across the entire amino acid sequence. If individual areas are selected, homologies of approximately 80% to 100% can be found in some sections (Al-Feel et _. Al, supra). The same organizational form of eukaryotic ACCs with regard to the sequence of the domains BC-BCCP-CT is remarkable. This speaks for an early fusion of the individual genes of the prokaryotes in the course of the evolution of the eukaryotes. The high level of conservation of the ACCs has been experimentally confirmed by the high cross-reactivity of antibodies between rats, chickens and yeast (Al-Feel et _. Al, supra).

The regulation of acetyl-CoA carboxylase in the various organisms is still largely unclear. So far, the enzyme activity in plants has been investigated on two different experimental systems, namely in chloroplasts and developing rapeseed. It has been found that in developing rapeseed, the activity of the ACC is induced before lipid storage, but then decreases rapidly when full lipid storage has been achieved (E. Turnham and DH Northcote (1983) Biochem. J. 212, S. 223-229). This means that regulation takes place via the end product. In addition, the ACC seems to regulate the entire de novo fatty acid biosynthesis as an enzyme that limits the turnover rate (PD Simcox, W. Garland, V. De Lica, DT Canvin and DT Dennis, Can J Bot 57, p. 1008- 1014 (1979); Turnham and Northcote, supra). These experimental findings make acetyl-CoA carboxylase in plants an interesting object for intervening in fatty acid metabolism with the perspective of increasing yield or changing the fatty acid pattern with a suitable overproduction of the ACC in the seed.

Studies on the mode of action of various herbicides used to control grasses, such as gramineae, in stands of dicotyledonous crops have shown that certain herbicides interfere with the metabolism of Gramineae by inhibiting the ACC. So far, substances of three different classes of substances have been described which develop a herbicidal action through interaction with the ACC. For example, derivatives of aryloxyphenoxypropionic acid (eg diclofop, fenoxaprop, fluazifop and haloxyfop) inhibit (K. Kobek et al, Z. Naturforschung 43c, pp. 47-54 (1988)), cyclohexane-1,3-dione (eg cycloxydim , Clethodim and Setoxydim) (M. Focke and HK Lichtenthaler, Z. Naturforschung 42c, pp. 1361-1363 (1987)) or of PP600 (3-isopropyl-6- (N- [2, 2-dimethylpropyl] -acetamido- 1, 3, 5-triazin-2, 4- (1H, 3H) dione) (KA Walker, SM Ridley and JL Lewis Harwood, Phytochem 29, pp. 3743-3747 (1990)) the ACC sensitive plants. Like the inhibitory The effect is in detail and why ACCs are not inhibited by dicotyledonous plants is currently still unclear.

EP-A-0 469 810 describes a biotin-containing polypeptide with a molecular weight of 50 kD, which is a subunit of a vegetable acetyl-CoA carboxylase. However, it has been found, among other things, that the 229 bp clone CC 8 in FIG. 8 has no amino acid reading frame which has meaningful homology to one of the known ACC amino acid sequences. This inevitably leads to the conclusion that the antibody used in EP-A-0 469 810 is not specific to ACC or at least one subunit of ACC.

It is an object of the invention to provide a DNA sequence with which, on the one hand, the quality and quantity of vegetable oils or fats can be changed by homologous or heterologous expression in plant systems and, on the other hand, by heterologous expression herbicide resistance in, for example, useful plants to a wide variety of crops Herbicides can be awarded or transferred. This object is achieved with a DNA sequence according to claim 1.

The invention further relates to genomic clones which contain a DNA sequence which codes for the acetyl-CoA carboxylase, and the alleles and derivatives of this DNA sequence.

The invention also relates to a process for the production of plants, plant parts and plant products, in which a DNA sequence which codes for the acetyl-CoA carboxylase is transmitted by genetic engineering.

Finally, the invention also relates to the use of this DNA sequence for conferring or transferring herbicide resistance or changing the quality and quantity of vegetable oils and fats.

The figures serve to explain the present invention. Show it:

FIG. 1 shows a sequence comparison of the amino acid sequences of biotin-dependent and related enzymes in their BC domain;

FIG. 2 shows the representation of the DNA or amino acid sequence of the degenerate oligonucleotides 3455 and 3464;

3a shows the DNA sequence and the one derived therefrom

Amino acid sequence in the one-letter code of the 260 bp PCR fragment as a specific hybrid locating probe;

Figure 3b the comparison of the amino acid sequence of the ACC

Rat (top line) with the amino acid sequence from FIG. 3a (bottom line);

Figure 4 shows the restriction maps in the genomic

Cloning BnACC3, BnACCδ, BnACCIO and BnACCl inserted DNA sequences;

FIG. 5 shows the DNA sequence of the acetyl-CoA carboxylase

Gene;

FIG. 6 shows the functional regions in the DNA sequence from FIG. 5 and the amino acid sequences derived from the DNA sequences in the one-letter code;

Figure 7 is a schematic representation of the functional

Regions of the DNA sequence from FIG. 6; and

FIG. 8 shows a Southern blot hybridization (cross hybridization) of different genomic plant DNA with part of the ACC gene of the genomic clone BnACC8.

It goes without saying that allelic variants and derivatives of the DNA sequence according to the invention are also covered in the context of the invention, provided that these modified DNA sequences code for acetyl-CoA carboxylase. The allelic variants and derivatives include, for example, deletions, substitutions, insertions, inversions or additions of the DNA sequence according to the invention. The gene for acetyl-CoA carboxylase is present in all plants and can therefore be isolated from them in various ways. For example, the gene can be isolated with the aid of oligonucleotide probes or specific antibodies from genomic plant DNA banks or its cDNA from cDNA banks. Rapeseed (Brassica napus) of the Akela variety has proven to be a particularly suitable plant material.

In the present invention, a genome of the rape genome (Brassica napus) of the Akela variety was used as a starting material for the isolation of genomic clones which contain the gene for the ACC, and was set up in a phage. This gene bank was searched for genes for the ACC using a hybridization probe produced by means of PCR (polymerase chain reaction). In this way, a genomic clone called BnACC8 was isolated, which contains the complete structural gene (protein coding region (exons and introns)) of the ACC from oilseed rape on a 13.7 kb Xbal fragment. This genomic clone is deposited under the number DSM 7384.

Furthermore, the genomic clones BnACC3, BnACCIO and BnACCl were isolated, which likewise contain the structural gene of the ACC from rapeseed or at least parts thereof on approximately 20 kb, 15 kb and 15 kb DNA fragments, respectively.

The 13.7 kb DNA fragment was subcloned in the form of Xbal / Smal fragments into suitable vectors and sequenced. The amino acid sequences derived from the DNA sequences were compared with the ACC amino acid sequence of the rat from FIG. 2 of the article by F. Lopez-Casillas, supra, by computer analysis. It has been found on the basis of amino acid sequence homologies that the 13.7 kb DNA fragment contains the acetyl-CoA carboxylase gene. In addition, an approximately 2 kb DNA fragment of the approximately 20 kb DNA sequence from BnACC3 was sequenced.

FIG. 4 shows the restriction maps of the DNA fragments inserted in the genomic clones BnACC3, BnACC8, BnACCIO and BnACCl. With the exception of the overlapping clones BnACC3 and BnACC8, which belong to a class of genes, only one representative (BnACCIO and BnACCl) of two further classes of genomic clones was shown. The areas marked in black indicate areas of DNA that hybridize with the probe used. The DNA fragments are delimited by the interfaces of the cloning vector Lambda FI __ "* TI, which are symbolized by" Y ": Xbal, Sacl, Notl, Sacl and Sall on one side or Sall, Sacl, Notl, Sacl and Xbal on the The sequenced areas of the 13.7 kb DNA fragment from BnACC8 and the approximately 2 kb DNA fragment from BnACC3 were marked by white bars.

11.9 kb of the 13.7 kb DNA fragment from BnACC8 were sequenced using conventional methods. Furthermore, approximately 2 kb were sequenced from the second Sall site of the approximately 20 kb DNA fragment from BnACC3 in the 3 'direction until overlapping with clone BnACCδ. Both clones overlap in the 5 'range. Sequence comparison results in a DNA sequence with a length of 13.753 kb.

The complete DNA sequence of the 13.753 kb from the 11.9 kb of the 13.7 kb DNA fragment from BnACC8 and 1904 bp of the approximately 20 kb DNA fragment from BnACC3 is shown in FIG. 5. The DNA sequence at the 5 'end comprises the sequence from BnACC3, starting from the second SalI site and extends at the 3' end with 678 bp beyond the EcoRI site of the BnACC8. The DNA sequence of the 11.9 kb of the 13.7 kb DNA fragment from BnACCδ begins at position 1905 in FIG. 5. The 13.753 kb DNA fragment contains the acetyl-CoA carboxylase structural gene and the promoter, the structural gene already being on the 11.9 kb of the 13.7 kb DNA fragment from BnACC8. For this purpose, reference is made to FIG. 6, which shows the DNA sequence from FIG. 5 with its functional areas. Regulatory elements such as the CAAT box (positions 2283-2286), the TATA box (positions 2416-2419) and a polyadenylation signal (positions 13284-12289) are underlined. The first approximately 600 bp from position 1905 (11.9 kb from BnACCδ) already form part of the promoter; the preceding 1904 bp contain the entire promoter and come from BnACC3. The ATG start codon of the ACC is in position 2506 and the associated TGA stop codon in position 13253. The exon / intron boundaries have been highlighted in black.

The corresponding amino acid sequences were given for the black highlighted exon regions in the sequence. The exon / intron limits were determined on the basis of the similarity to acetyl-CoA carboxylases from other organisms (rat) (F. Lopez-Casillas, supra), unless these limits have been determined by means of PCR. The first exon of the gene begins at "ATG" as the start codon with an open reading frame. The 5 'non-translated areas are therefore not highlighted in black. The marking of the last exon ends at the corresponding stop codon, so that the 3 'untranslated region is also not marked.

The functional regions of the ACC sequence of the clone BnACCδ are shown schematically in FIG. The exons are highlighted in black. ATG means the start codon, MKM the conserved biotin binding site and TGA the stop codon. Furthermore, the three domains can be clearly assigned to the sequence: BC = biotin carboxylase. BCCP = biotin carboxy carrier protein, CT = carboxyl transferase. The three domains were due to homology to the ACC of other organisms.

In cross hybridizations with genomic DNA from Arabidopsis thaliana, Brassica napus, Avena sativa, Hordeum vulgare, Oryza sativa, Triticum aestivum and Zea mays, it was found that parts of the ACC sequence according to the invention are suitable for isolating ACC genes from other plants. In Figure δ a Smal / Sacl fragment (see Figure 4) from the clone BnACCδ was used as the DNA probe. The genomic DNAs of the different plants were digested with Eco RI and subjected to a Southern blot. The cross reactivity of the probe is observed in mono- and dicotyledonous plants.

The DNA sequence according to the invention, which codes for the acetyl-CoA carboxylase, the alleles and derivatives of this DNA sequence can be introduced or transferred into plants for the regulation of the fatty acid metabolism (in the form of anti-sense or overexpression) with the aid of genetic engineering methods .

Antisense constructions e.g. with sequences from positions 1905 to 3187, 318δ to δlOδ and 11039 to 12646 of the DNA sequence according to the invention from FIG. 6 can be used to inhibit the activity of the ACC in a plant. This can be done in particular by checking fragments of the ACC gene by checking fragments of the ACC gene by means of suitable regulatory elements (promoters) in the seed. This can cause a build-up of acetyl-CoA, since this intermediate can no longer flow into the fatty acid metabolism and thus the metabolism e.g. a plant cell influences:

1. With suitable regulatory elements, a "suicide gene" can thus be produced if an antisense construction leads to the formation of fatty acids in a Cell is omitted. In the fight against plant diseases, a hypersensitive reaction can be triggered in this way.

2. By adding additional genes whose gene products use acetyl-CoA, the backflow of acetyl-CoA can be prevented. For example, the genes for the synthesis of, for example, polyhydroxybutyrate (PHB) (Piorier et _. Al. 1992, Science 256, pages 520-523) can be stored in particular tissues / organs / cell types of a plant, preferably storage tissues such as seeds ( Endosperm, cotyledon); Root; various types of tubers) can be expressed. If an ACC antisense construction is simultaneously expressed in the same parts of the plant, the unused acetyl-CoA can be used for the synthesis of PHBs.

Oligonucleotides can be derived from the DNA sequence according to the invention in order to synthesize a cDNA or pieces of a cDNA. This cDNA or pieces thereof can be used alone or in conjunction with parts of the genomic clone to isolate a complete cDNA. These cDNA or cDNA pieces can also be used for an antisense expression.

For example, individual cDNA fragments or the entire cDNA can be used to complement ACC mutants, for example in microorganisms. This means that the microorganisms (mutants from E. coli fabE; Silbert et. Al. 1976, J. Bacteriol. 126, pages 1351-1354); Harder et al. 1972, PNAS 69, pages 3105-3109 and from yeast (Schweizer et _. Al. About i960) under non-permissive conditions functionally complemented by the vegetable ACC and are directly dependent on the vegetable enzyme. This results in the possibility of selection for the vegetable enzyme and a test system for the development and optimization of inhibitors for the ACC. This allows next to better active ingredients for use as herbicides, resistant forms of the ACC enzyme after mutagenesis of the gene (or regions of the gene) can be developed or selected.

The cDNA can also be used to recover larger amounts of the protein or parts of the protein. This manufactured protein can be used for studies on the reaction mechanism and the regulation or to elucidate the three-dimensional structure of the enzyme or parts of the enzyme. The latter point is particularly important for a "protein modeiling", since it allows the adaptation of e.g. Inhibitors allowed in the structure of the protein.

The ACC gene sequence, the alleles and derivatives of this sequence are preferably introduced into the plants together with suitable promoters, in particular in recombinant vectors.

All types of plants can be transformed for this purpose. In this context, useful plants, garden plants and ornamental plants may be mentioned. Brassica napus, B. rapa, coconut and oil palm, sunflower and flax are particularly preferred among the useful plants.

The DNA sequence according to the invention, which codes for the ACC, can be used in particular to achieve herbicide resistance in useful plants, including in particular cereal plants, against certain herbicides. Maize, wheat, barley, rice and rye can be mentioned as preferred plants to be transformed.

The genetic engineering introduction of the ACC-DNA sequence, the alleles and derivatives of this sequence can be carried out using conventional transformation techniques. Such techniques include methods such as direct gene transfer, such as microinjection, electroporation, particle gun, viral Vectors and liposome-mediated transfer as well as the transfer of corresponding recombinant Ti plasmids or Ri plasmids and the transformation by plant viruses.

In the case of a cell culture of a monocotyledonous plant, such as barley, wheat or maize, the detection of the transformation can be carried out by selection with a suitable herbicide. Furthermore, the detection can be carried out by Southern blots with, for example, intron sequences of the rapeseed ACC DNA as a hybridization probe.

The invention thus also relates to plants, plant parts and plant products which have been produced or transformed by one of the above processes.

The following examples serve to explain the invention.

Examples

Example 1: Preparation of the Hybridization Probe for Acetyl-CoA Carboxylase (ACC)

(a) Preparation of degenerate oligonucleotides

Based on a sequence comparison of different biotin-containing proteins, synthetic oligonucleotides were derived from conserved sections of the ACC sequences. For this purpose, reference is made to FIG. 1, which shows a sequence comparison of the amino acid sequence of biotin-dependent and related enzymes in their BC domain. This figure goes to Figure 3 from the publication by Kondo et _. al, supra, back. The abbreviations in the left column have the following meanings: EACC = ACC from E. coli; cACC = ACC from chicken; rPCCof = α subunit of the propionyl-CoA carboxylase from the

Rat; yPC = yeast pyruvate carboxylase and ECPSN = n-terminal half of the carbamoyl phosphate synthetase.

Identical amino acids are framed and strictly preserved residues are marked with dots. In addition, the conserved sequences in FIG. 1 which served for the production or derivation of the degenerate oligonucleotides 3455 and 3464 were highlighted by arrows and numbers.

The oligonucleotides were synthesized on an Applied Biosystems DNA synthesizer (Model 380B) and are shown in FIG. 2. Both oligonucleotides are shown in 5 '-3' orientation, so that when comparing with FIG. 1, the amino acid sequence of oligonucleotide 3464 must be read in the opposite direction.

Due to the degenerate genetic code and the possible variability of the amino acid sequence at individual positions, different bases were built into the oligonucleotides, e.g. C or T or A or G in oligonucleotide 3464. In addition, I was introduced, which can interact with all nucleotides and is therefore to be regarded as a non-specific base.

(b) Polymerase chain reaction (PCR)

Starting from one μg of polyA + RNA, Avian myeloblostosis virus (AMV) was used to reverse transcriptase for 30 minutes at a temperature of. 37 C performed a cDNA synthesis with the oligonucleotide 3464 as a primer. After inactivity Reverse transcriptase by heating for five minutes at a temperature of 95 C, the PCR reaction with 50 pmol final concentration per primer (3455 and 3464) and four units of Ampli-Taςp ^ polymerase (Perkin Elmer Cetus) was carried out in the same reaction mixture . The reactions were carried out under the following conditions:

a) Buffer conditions: 10 mM Tris-HCl, pH 8.0; 50 mM KCl; 1.5 mM MgCl ₂ ; 0.01% gelatin and 5 mM dNTPs;

b) Reaction temperatures: 3 minutes at a temperature of 92 C for the first denaturation, then 30 temperature cycles with 2 minutes each at a temperature of 92 C for the denaturation, 2 minutes at a temperature of 51 C for annealing the oligonucleotides and 2.5 minutes a temperature of 72 ° C for amplification of the DNA and finally 2.5 minutes at a temperature of 72 ° C to achieve a complete synthesis of the last synthesis products.

c) Cloning the amplification products

Protruding single-stranded DNA of the PCR products were filled in using Klenow polymerase (Sambrook et. Al, Molecular Cloning - A Laboratory Manual, 2nd ed. Cold Spring Harbor Laboratory Press, New York (1989)) and then phosphorylated with polynucleotide kinase (Sambrook et al, supra). The PCR products were purified according to standard protocols according to Sambrook et al, supra, by agarose gel electrophoresis, gel elution, purification with phenol / chloroform and the final precipitation with isopropanol. The on this

Purified DNA was cleaved into S al pBluescipt _§) -

Vector DNA ligated and cloned. d) DNA sequencing

For the determination of the DNA sequence of subclones produced in pBluescripc and those DNA sequences in which deletions were produced by means of exonuclease III (see also Example 2) (Sambrook et. Al, supra) were carried out according to the Sanger method et _. al, Proc. Natl. Acad. Be. 74, pp. 5463-5467 (1977). The sequence data were analyzed using the computer software of the "University of Wisconsin Genetics Computer Group" (Devereux et. Al, Nucl. Acids Res. 12, pp. 367-395 (1964)). The homology studies were carried out with the "Bestfit" program.

e) synthesis of a specific hybridization probe by means of PCR

Starting from polyA-RNA from immature rapeseed seeds (Brassica napus) (about 2-3 weeks old), DNA fragments were amplified by means of PCR reactions after a first-strand cDNA synthesis. The oligonucleotides required for this were synthesized on the basis of a homology comparison inter alia between the ACC from the chicken and from E. coli (FIG. 1). Mathematically, these (degenerate) oligonucleotides should be able to amplify a product of 260 bp length, which codes for 86 amino acids. Amplification products of this magnitude were therefore isolated from the product mixture obtained, cloned in pBluescripr- ^ and identified by DNA sequencing. In addition to other non-specific PCR products, one could be cloned that has the expected size of 260 bp and has an open reading frame of 86 amino acids (FIG. 3a). This product, if the homology is also included in the area of the oligonucleotides, has 77.9% identical amino acids compared to the ACC of the rat and a homology of 8δ, 4% (FIG. 3b). If one calculates the identity or homology only on the amplified sequence, i.e. without the In the area of the oligonucleotides that also allow mismatches, there are still values of 73.2% identical amino acids or 85.9% homology between the protein sequences of the ACC of rats and rapeseed. These numbers show that the cloned PCR product encodes part of the ACC from rapeseed and can thus be used as a specific hybridization probe. Due to the position of the homology to the ACC ACC of the rat between the amino acids of positions 304 and 389 (FIG. 3b), it was to be expected that the cloned PCR fragment would only recognize cDNAs which are over 6000 bp long.

Example 2: Characterization of a genomic clone with a DNA sequence coding for ACC

10 genomic clones were isolated and characterized from a gene bank of rape (Brassica napus) of the Akela variety, which had been constructed in the vector Lambda FlJrll (Stratagene), with the aid of the PCR fragment cloned as described in Example 2. Because of their restriction maps, they can be divided into 3 classes. FIG. 4 shows the restriction maps of the genomic clones BnACC3, BnACCδ, BnACCIO and BnACCl. The clone, BnACCδ, which belongs to the most frequently represented class, contains a DNA fragment with a size of 13.7 kb. This DNA fragment comprises the complete structural gene of the ACC from rapeseed. The DNA fragment was in the form of Xbal

Smal fragments in pBluescript- ^ subcloned sequenced. Furthermore, an approximately 3.2 kb Sall-Smal fragment DNA fragment of the approximately 20 kb DNA fragment from BnACC3 was subcloned and approximately 2 kb of which was sequenced in 3 'direction from the Sall interface.

Claims

1. DNA sequence coding for the acetyl-CoA carboxylase and the alleles and derivatives of this DNA sequence.

2. DNA sequence according to claim 1, d a d u r c h g e k e n n z e i c h n e t that it is isolated from plants.

3. DNA sequence according to claim 2, d a d u r c h g e k e n n z e i c h n e t that it is isolated from oilseed rape (Brassica napus).

4. DNA sequence according to one of claims 1 to 3, characterized in that it has a size of 13.7 kb and contains the complete structural gene and at least parts of the promoter of acetyl-CoA carboxylase.

5. Genomic clone which contains a DNA sequence which codes for the acetyl-CoA carboxylase, and the alleles and derivatives of this DNA sequence.

6. Genomic clone according to claim 5, d a d u r c h g e k e n n z e i c h n e t that the DNA sequence is isolated from plants.

7. Genomic clone according to claim 6, that the DNA sequence from oilseed rape (Brassica napus) is isolated.

8. Genomic clone according to one of claims 5 to 7, that the DNA sequence has a size of 13.7 kb and contains the complete structural gene and at least parts of the promoter of the acetyl-CoA carboxylase.

9. Genomic clone BnACCδ (DSM 7384).

10. Genomic clones BnACCl, BnACC3 and BnACCIO, which contain the DNA sequence for the complete structural gene of acetyl-CoA carboxylase or at least parts thereof and the alleles and derivatives of this DNA sequence.

11. Genomic clones according to claim 10, so that the DNA sequence for the structural gene is isolated from plants.

12. Genomic clones according to claim 11, characterized in that the DNA sequence for the structural gene from rape (Brassica napus) is isolated.

13. Genomic clones according to claim 12, so that at least the genomic clone BnACC3 contains the DNA sequences for the promoter for the structural gene of the acetyl-CoA carboxylase.

14. DNA sequence which contains the structural gene and the promoter for the acetyl-CoA carboxylase according to FIG. 5.

15. A process for the production of plants, parts of plants and plant products which have herbicide resistance, in which a DNA sequence according to one of claims 1 to 4 or claim 14 or a DNA sequence derived from the genomic clones according to one of claims 5 to 13 genetic engineering path is transferred.

16. A process for the production of plants, plant parts and plant products, the quality and quantity of which is changed with regard to oleic and fatty acid production, in which a DNA sequence according to one of claims 1 to 4 or claim 14 or one of the genomic clones according to one of the Claims 5 to 13 derived DNA sequence is transferred by genetic engineering.

17. The method according to claim 14 or 15, characterized in that the DNA sequence is transferred by microinjection, electroporation, particle gun, transfer of corresponding recombinant Ti plasmids or Ri plasmids, liposome-mediated transfer or by plant viruses.

18. Use of a DNA sequence according to one of claims 1 to 4 or claim 14 or a DNA sequence derived from the genomic clones according to one of claims 5 to 13 for the transmission of herbicide resistance in plants.

19. Use of a DNA sequence according to one of claims 1 to 4 or claim 14 or a DNA sequence derived from the genomic clones according to one of claims 5 to 13 for changing the quality and quantity with regard to the oil and fatty acid production in plants.

20. Plants, parts of plants and plant products produced by a process of claim 15 or 16 and 17.