Expression and purification of recombinant neurotensin in Escherichia coli.

Journal article

An expression system has been designed for the rapid and economic expression of recombinant neurotensin for biophysical studies. A synthetic gene for neurotensin (Glu(1)-Leu(2)-Tyr(3)-Glu(4)-Asn(5)-Lys(6)-Pro(7)-Arg(8)-Arg(9)-Pro(1 0)-Tyr(11)-Ile(12)-Leu(13)) was cloned into the pGEX-5X-2 vector to allow expression of neurotensin as a glutathione S-transferase (GST) fusion protein. The inclusion of a methionine residue between the glutathione S-transferase and the neurotensin has facilitated the rapid cleavage of the neurotensin from its carrier protein. Purification of recombinant neurotensin was performed by reverse-phase HPLC. This method produced a relatively high yield of peptide and offers the potential for economic partial or uniform labeling of small peptides (<15 amino acids) with isotopes for NMR or other biophysical techniques.

Authors: Williamson, P; Roth, J; Haddingham, T; Watts, A

• Publication status: Published
• Journal: Protein expression and purification
• Volume: 19
• Issue: 2
• Pages: 271-275
• Publication date: 2000-07-01
• DOI: 10.1006/prep.2000.1246
• EISSN: 1096-0279
• ISSN: 1046-5928
• Language: English
• Keywords: Glutathione Transferase; Chromatography, High Pressure Liquid; Escherichia coli; Recombinant Fusion Proteins; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Electrophoresis, Polyacrylamide Gel; Neurotensin