Preparation of polypeptides having an amide carboxyl terminal en

Details Preparation of polypeptides having an amide carboxyl terminal en Preparation of polypeptides having an amide carboxyl terminal en

1994-03-04

1995-10-10

Fox, David T.

Chemistry: molecular biology and microbiology

Micro-organism, tissue cell culture or enzyme using process...

Recombinant dna technique included in method of making a...

435 681, 435 694, 435 698, 435 712, 4351723, 4352523, 43525233, 4353201, 536 234, 536 2351, C12N 1516, C12N 1562, C12N 1570, C12P 2106

Patent

active

054570336

ABSTRACT:
Polypeptides having a carboxamide carboxyl terminal end and a methionine residue, which is optionally bonded to a bacterial protein, on the amino end can be prepared by synthesis by genetic engineering methods of the corresponding polypeptide having at the C terminal end a glycine residue, and conversion of the latter enzymatically into the amino group of the desired carboxamide group. Peptides which have the aminoacid sequence of growth hormone releasing factor, part sequences thereof, or modifications of these peptides, are readily accessible by this means. The synthesis by genetic engineering methods is advantageously carried out via two gene fragments which are synthesized chemically from smaller, single-stranded structural units. The two gene fragments are then linked enzymatically to give the complete gene, which is incorporated into a suitable vector, amplified there, and the peptide is isolated directly or as a fused protein, and is converted enzymatically into the desired amide.

REFERENCES:
Guillemin et al., Science, 218: 585-587 (1982).
Bradbury et al., Nature, 298: 686-688 (1982).
Husain et al., FEBS Letters, 152: 277-281 (1983).
Gait et al., Nucleic Acids Res., 8: 1081-1096 (1980).
Amann et al., Gene, 25: 167-178 (1983).
Gubler et al. 1983. Proc. Natl. Acad. Sci. USA 80:4311-4314.
Bradbury et al. 1982. Nature 298: 686-688.
Smith et al. 1982. Nucleic Acids Research 10(150) 4467-4482.

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