Plant acetyl-coa carboxylase polypeptide and gene

Claim 2

The polypeptide of claim 1 wherein said polypeptide contains amino acids encoded by the cDNA sequence shown in Figure 8.

Claim 3

The polypeptide of claim 1 wherein said polypeptide catalyzes the carboxylation of acetyl-CoA to form malonyl-CoA.

Claim 4

A polypeptide derived from the polypeptide of claim 3, wherein said polypeptide catalyzes the carboxylation of acetyl-CoA to form malonyl-CoA.

Claim 6

The method of claim 5 wherein said tetrameric avidin to streptavidin bound to a solid support is in a chromatographic column.

Claim 7

The method of claim 5 further comprising the step of separating all recovered biotin-containing polypeptides by molecular weight, thereby recovering said polypeptides in purified form.

Claim 8

A method of obtaining the polypeptide of claim 1 comprising the steps of: freezing plant tissue containing acetyl-CoA carboxylase by contacting said tissue with liquid nitrogen; pulverizing said tissue while in contact with said liquid nitrogen; homogenizing said pulverized tissue to form a homogenate containing said polypeptide; extracting said homogenate with a protein-extracting solution to form an extract containing said polypeptide; separating said extract from the remaining part of said homogenate; contacting said separated extract with tetrameric avidin or tetrameric streptavidin bound to a solid support for a sufficient time and under appropriate conditions to bind said polypeptide to said avidin or streptavidin; removing said polypeptide from said avidin or streptavidin; separating all recovered biotin-containing polypeptides by molecular weight; and recovering in purified form the polypeptide with a molecular weight of about 50kD.

Claim 9

A method of obtaining plant acetyl-CoA carboxylase in purified form comprising the steps of: extracting plant tissue containing acetyl-CoA carboxylase with a protein-extracting solution to form an extract containing said plant acetyl-CoA carboxylase; contacting said extract with antibodies to the polypeptide of claim 1 for a sufficient time and under appropriate conditions to bind said acetyl-CoA carboxylase to said antibodies; separating said acetyl-CoA carboxylase bound to said antibodies from said extract; and removing said acetyl-CoA carboxylase from said antibodies, thereby recovering said acetyl-CoA carboxylase in purified form.

Claim 10

The method of claim 9 wherein said antibodies are bound to a solid support.

Claim 11

The method of claim 9 wherein said separating step comprises contacting said extract containing the complex comprising acetyl-CoA carboxylase bound to said antibodies with an antibody-binding compound bound to a solid support for a sufficient time and under appropriate conditions to bind said complex to said antibody-binding compound.

Claim 13

The method of claim 12 further comprising the step of recovering said cDNA sequence from said clone.

Claim 14

The method of claim 13 further comprising the step of evaluating said cDNA sequence to confirm that it codes for said polypeptide.

Claim 15

The said purified cDNA sequence obtainable by the method of claim 13.

Claim 16

An isolated or substantially purified DNA sequence that codes for the polypeptide of claim 1.

Claim 17

The DNA sequence of claim 16 wherein said DNA is cDNA.

Claim 18

The DNA sequence of claim 17 wherein said cDNA sequence contains the nucleotide sequence shown in Figure 8.

Claim 19

An isolated or substantially purified DNA sequence derived from the DNA sequence of claim 18 by single or multiple mutations.

Claim 20

A DNA sequence that hybridizes with the cDNA sequence of claim 18 under conditions of high stringency.

Claim 21

A recombinant DNA sequence comprising the DNA sequence of claim 16 operably linked to appropriate regulatory control nucleic acid sequences that are capable of effecting the expression of said DNA sequence in transformed host cells.

Claim 22

An expression vector for expressing DNA that codes for the protein of claim 1 in a compatible host comprising an expression vector capable of transforming a procaryotic or eucaryotic cell and the DNA of claim 16 inserted into said vector in proper orientation and correct reading frame for expression.

Claim 23

A host cell transformed with the recombinant DNA sequence of claim 21.

Claim 24

The host cell of claim 23 wherein said cell is a plant cell.

Claim 25

The host cell of claim 24 wherein said plant cell is a monocot cell.

Claim 26

The recombinant protein obtainable from the transformed cell of claim 23.

Claim 27

A method for producting the protein of claim 1 comprising the steps of: culturing host cells transformed by a recombinant DNA sequence comprising a DNA sequence that codes for the protein of claim 1 operably linked to appropriate regulatory control nucleic acid sequences that are capable of effecting the expression of said DNA sequence in said transformed cells; and recovering the polypeptide whose expression has been coded for by said sequence.