A general method for retrieving the components of a genetically engineered fusion protein.
Escherichia coli expression vectors encoding an acid-labile aspartyl-proline (Asp-Pro) dipeptide bridging two protein sequences were constructed and used to synthesize two different bovine growth hormone (bGH) fusion proteins. The codons GAT-CCX coding for Asp-Pro are provided by the recognition site for Bam HI (GGATCC). Treatment of the bGH fusion proteins at low pH in the presence of guanidine hydrochloride releases the bGH moiety from the fusion protein. The release of the bGH from the fusion protein specifically requires the Asp-Pro dipeptide linking the bGH sequence to the fusion protein. The bGH released from the fusion protein retains anti-bGH immunoreactivity as well as the ability to bind to growth hormone receptor in vitro.
D000143:Acids; D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D004262:DNA Restriction Enzymes / Q000378:metabolism; D015245:Deoxyribonuclease BamHI; D004926:Escherichia coli / Q000235:genetics; D005818:Genetic Engineering / Q000379:methods*; D013006:Growth Hormone / Q000235:genetics*; D006868:Hydrolysis; D011956:Receptors, Cell Surface / Q000378:metabolism; D011986:Receptors, Somatotropin; D011994:Recombinant Proteins / Q000235:genetics*; D014364:Tryptophan / Q000235:genetics
D000143:Acids; D011956:Receptors, Cell Surface; D011986:Receptors, Somatotropin; D011994:Recombinant Proteins; D014364:Tryptophan; D013006:Growth Hormone; D004262:DNA Restriction Enzymes; D015245:Deoxyribonuclease BamHI