DNA for expression of bovine growth hormone
OwnerHerman A. de Boer, Herbert L. Heyneker +1inventors
InventorsHerman A. de Boer, Herbert L. Heyneker, Peter H. Seeburg
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Abstract
High-level expression of bovine growth hormone is achieved using DNA encoding bovine growth hormone which, when transcribed, produces messenger RNA (mRNA) that at its 5' end is substantially free of secondary structure that is capable of interfering with translation of the mRNA.
Claims6
An isolated DNA comprising a first nucleotide sequence which encodes a polypeptide comprising the bovine growth hormone N-terminal sequence ##STR5## and a second nucleotide sequence which encodes the bovine growth hormone C-terminal amino acid sequence ##STR6## wherein the first and second nucleotide sequences are covalently joined to encode the complete amino acid sequence of a BGH, and wherein the first nucleotide sequence is selected so as to provide, upon transcription, a corresponding messenger RNA sequence that is free of secondary structure having a thermodynamic energy arithmetically less than or equal to the thermodynamic energy of the secondary structure formed by homologous base pairing between nucleotides 46 to 51 and nucleotides 73 to 78 of the natural messenger RNA encoding BGH.
A microorganism containing a DNA that comprises a first nucleotide sequence which encodes a polypeptide comprising the bovine growth hormone N-terminal sequence
- __________________________________________________________________________ Phe Pro Ala Met Ser Leu Ser Gly Leu Phe Ala Asn Ala Val Leu Arg Ala Gln His Leu His Gln Leu Ala Ala __________________________________________________________________________
- and a second nucleotide sequence which encodes the bovine growth hormone C-terminal amino acid sequence
- __________________________________________________________________________ Asp Thr Phe Lys Glu Phe Glu Arg Thr Tyr Ile Pro Glu Gly Gln Arg Tyr Ser Ile Gln Asn Thr Gln Val Ala Phe Cys Phe Ser Glu Thr Ile Pro Ala Pro Thr Gly Lys Asp Glu Ala Gln Gln Lys Ser Asp Leu Glu Leu Leu Arg Ile Ser Leu Leu Leu Ile Gln Ser Trp Leu Gly Pro Leu Gln Phe Leu Ser Arg Val Phe Thr Asn Ser Leu Val Phe Gly Thr Ser Asp Arg Val Tyr Glu Lys Leu Lys Asp Leu Glu Glu Gly Ile Leu Ala Leu Met Arg Glu Leu Glu Asp Gly Thr Pro Arg Ala Gly Gln Ile Leu Lys Gln Thr Tyr Asp Lys Phe Asp Thr Asn Met Arg Ser Asp Asp Ala Leu Leu Lys Asn Tyr Gly Leu Leu Ser Cys Phe Arg Lys Asp Leu His Lys Thr Glu Thr Tyr Leu Arg Val Met Lys Cys Arg Arg Phe Gly Glu Ala Ser Cys Ala Phe __________________________________________________________________________
- wherein the first and second nucleotide sequences are covalently joined to encode the complete amino acid sequence of a BGH, and wherein the first nucleotide sequence is selected so as to provide, upon transcription, a corresponding messenger RNA sequence that is free of secondary structure having a thermodynamic energy arithmetically less than or equal to the thermodynamic energy of the secondary structure formed by homologous base pairing between nucleotides 46 to 51 and nucleotides 73 to 78 of the natural messenger RNA encoding BGH.
A microorganism containing a DNA that is operably linked to a promoter which controls expression of the DNA in the microorganism, which DNA comprises a first nucleotide sequence which encodes a polypeptide comprising the bovine growth hormone N-terminal sequence
- __________________________________________________________________________ Phe Pro Ala Met Ser Leu Ser Gly Leu Phe Ala Asn Ala Val Leu Arg Ala Gln His Leu His Gln Leu Ala Ala __________________________________________________________________________
- and a second nucleotide sequence which encodes the bovine growth hormone C-terminal amino acid sequence
- __________________________________________________________________________ Asp Thr Phe Lys Glu Phe Glu Arg Thr Tyr Ile Pro Glu Gly Gln Arg Tyr Ser Ile Gln Asn Thr Gln Val Ala Phe Cys Phe Ser Glu Thr Ile Pro Ala Pro Thr Gly Lys Asp Glu Ala Gln Gln Lys Ser Asp Leu Glu Leu Leu Arg Ile Ser Leu Leu Leu Ile Gln Ser Trp Leu Gly Pro Leu Gln Phe Leu Ser Arg Val Phe Thr Asn Ser Leu Val Phe Gly Thr Ser Asp Arg Val Tyr Glu Lys Leu Lys Asp Leu Glu Glu Gly Ile Leu Ala Leu Met Arg Glu Leu Glu Asp Gly Thr Pro Arg Ala Gly Gln Ile Leu Lys Gln Thr Tyr Asp Lys Phe Asp Thr Asn Met Arg Ser Asp Asp Ala Leu Leu Lys Asn Tyr Gly Leu Leu Ser Cys Phe Arg Lys Asp Leu His Lys Thr Glu Thr Tyr Leu Arg Val Met Lys Cys Arg Arg Phe Gly Glu Ala Ser Cys Ala Phe __________________________________________________________________________
- wherein the first and second nucleotide sequences are covalently joined to encode the complete amino acid sequence of a BGH, and wherein the first nucleotide sequence is selected so as to provide, upon transcription, a corresponding messenger RNA sequence that is free of secondary structure having a thermodynamic energy arithmetically less than or equal to the thermodynamic energy of the secondary structure formed by homologous base pairing between nucleotides 46 to 51 and nucleotides 73 to 78 of the natural messenger RNA encoding BGH.
Dependencies are inferred from the claim text.