Chemoenzymatic Gene Synthesis of A Gene for Human Growth Hormone Releasing Hormone (hGHRH), its Expression in E. Coli and Enzymatic Amidation

Joachim W. Engels Universität Frankfurt, Institut für Organische Chemie und Biologische Chemie, D-6000, Frankfurt/Main, 50

Jan Glauder Universität Frankfurt, Institut für Organische Chemie und Biologische Chemie, D-6000, Frankfurt/Main, 50

Hubert Mülliner Hoechst AG, P.O. box 800320, D-6230, Frankfurt/Main, 80

Eugen Uhlmann Hoechst AG, P.O. box 800320, D-6230, Frankfurt/Main, 80

Waldemar Wetekam Hoechst AG, P.O. box 800320, D-6230, Frankfurt/Main, 80

T. Hashimoto Gotoh Hoechst AG, P.O. box 800320, D-6230, Frankfurt/Main, 80

Barbara Scheikl-lenz Universität Frankfurt, Institut für Organische Chemie und Biologische Chemie, D-6000, Frankfurt/Main, 50

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Abstract

A gene coding for hGHRH was constructed from chemically synthesized oligonucleotides in two fragments; After subcloning and joining the total gene was expressed in E. coli as a fusion protein with β-galactosidase. Chemical cleavage thereof with cyanogen bromide, purification of Leu²⁷ GHRH-Gly⁴⁵ by chromatography followed by enzymatic conversion yielded Leu²⁷ GHRHNH₂.

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Chemoenzymatic Gene Synthesis of A Gene for Human Growth Hormone Releasing Hormone (hGHRH), its Expression in E. Coli and Enzymatic Amidation

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Chemoenzymatic Gene Synthesis of A Gene for Human Growth Hormone Releasing Hormone (hGHRH), its Expression in E. Coli and Enzymatic Amidation

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