Production of proteins in procaryotes
AssigneeMonsanto Company
InventorGwen G. Krivi
Life of this patent
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Abstract
A method for preparing polypeptides in bacteria with an alanine rather than a methionine at the N-terminus. The DNA sequence expressed has an alanine codon immediately following from one to about three contiguous methionine codons including a translation start signal and allows for the expression of polypeptides having the amino acid sequence of, for example, naturally occurring eucaryotic proteins such as various bovine and porcine somatotropin species.
Claims29
A method for bacterially producing a mature bovine growth hormone polypeptide having an N-terminus alanine which comprises:
- causing expression of heterologous DNA in selected bacteria, said DNA containing the codons for said polypeptide and having the codon for the N-terminus alanine immediately downstream of a translation start signal/methionine codon,
- the expression resulting in producing within said selected bacteria said mature bovine growth hormone polypeptide having an N-terminus alanine;
- and
- recovering the polypeptide having an N-terminus alanine produced within said bacteria.
A method for producing in a procaryotic organism a mature heterologous bovine growth hormone polypeptide having an N-terminus alanine which comprises:
- selecting a DNA coding sequence for said polypeptide,
- inserting said coding sequence into an expression vector to obtain a gene sequence comprising a promoter operative in said procaryotic organism, a ribosome binding site, a translation start signal comprising a methionine codon immediately upstream of the DNA coding sequence, and a translation stop codon immediately downstream of said DNA Coding sequence,
- introducing said expression vector carrying said gene sequence into said procaryotic organism,
- culturing said organism under conditions which allow for expression of said DNA and production of said polypeptide, and recovering said polypeptide having an N-terminus alanine from said procaryotic organism.
A method for bacterially producing a mature porcine growth hormone polypeptide having an N-terminus alanine which comprises:
- causing expression of heterologous DNA in selected bacteria, said DNA containing the codons for said polypeptide and having the codon for the N-terminus alanine immediately downstream of a translation start signal/methionine codon,
- the expression resulting in producing within said selected bacteria said mature porcine growth hormone polypeptide having the N-terminus alanine;
- and
- recovering the polypeptide having the N-terminus alanine produced within said bacteria.
A method for producing in a procaryotic organism a mature heterologous porcine growth hormone polypeptide having a N-terminus alanine which comprises:
- selecting a DNA coding sequence for said polypeptide,
- inserting said coding sequence into an expression vector to obtain a gene sequence comprising a promoter operative in said procaryotic organism, a ribosome binding site, a translation start signal comprising a methionine codon immediately upstream of the DNA coding sequence, and a translation stop codon immediately downstream of said DNA coding sequence,
- introducing said expression vector carrying' said gene sequence into said procaryotic organism,
- culturing said organism under conditions which allow for expression of said DNA and production of said polypeptide, and recovering said polypeptide having a N-terminus alanine from said procaryotic organism.
Dependencies are inferred from the claim text.