Cloning, expression and purification of feline proinsulin.
Feline proinsulin was cloned and expressed using a bacterial expression system. It was then purified from inclusion bodies using size exclusion chromatography and further processed including reduction of the protein. Following refolding, proinsulin was purified by reversed-phase high-performance liquid chromatography (RP-HPLC). RP-HPLC and mass spectrometric analysis indicated that the proinsulin contained the correct disulfide bridging pattern. This proinsulin can be used for therapeutic and diagnostic purposes.
Hoenig
M
M
Department of Physiology and Pharmacology, College of Veterinary Medicine, The University of Georgia, Athens, GA 30602-7389, USA.
Caffall
Z F
ZF
McGraw
R A
RA
Ferguson
D C
DC
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002415:Cats / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular; D008969:Molecular Sequence Data; D010957:Plasmids; D011384:Proinsulin / Q000737:chemistry / Q000235:genetics*; D017510:Protein Folding; D012313:RNA / Q000737:chemistry / Q000235:genetics; D011994:Recombinant Proteins; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA
D011994:Recombinant Proteins; D012313:RNA; D011384:Proinsulin