Expression of atrial natriuretic factor as a cleavable fusion protein with chloramphenicol acetyltransferase in Escherichia coli.
Recombinant fusion proteins containing human atrial natriuretic factor, ANF(1-28) joined to chloramphenicol acetyltransferase (CAT) via cleavable linker sequences have been produced in Escherichia coli. The linker sequences were designed to allow the release of authentic ANF(1-28) following proteolytic cleavage by enterokinase or thrombin, or chemical cleavage with 2-(2-nitrophenylsulphenyl)-3-methyl-3'-bromoindolenine. Proteins, containing ANF(1-28) fused to the carboxyl-terminal region of CAT (using the ScaI restriction site in the cat gene), were largely soluble in E. coli and were obtained in higher yield than analogues containing ANF(1-28) linked to shorter CAT sequences. The longer derivatives also retained CAT activity allowing subsequent purification by affinity chromatography.
Dykes
C W
CW
Genetics Unit, Glaxo Group Research Limited, Greenford, England.
Bookless
A B
AB
Coomber
B A
BA
Noble
S A
SA
Humber
D C
DC
Hobden
A N
AN
D000123:Acetyltransferases / Q000302:isolation & purification*; D000595:Amino Acid Sequence; D009320:Atrial Natriuretic Factor / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D001483:Base Sequence; D015500:Chloramphenicol O-Acetyltransferase; D004765:Enteropeptidase / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology* / Q000378:metabolism; D005786:Gene Expression Regulation; D005822:Genetic Vectors; D006868:Hydrolysis; D008969:Molecular Sequence Data; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification*; D011994:Recombinant Proteins / Q000302:isolation & purification*; D012862:Skatole / Q000031:analogs & derivatives; D013917:Thrombin / Q000378:metabolism; D014158:Transcription, Genetic
D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; C005904:BNPS-skatole; D009320:Atrial Natriuretic Factor; D012862:Skatole; D000123:Acetyltransferases; D015500:Chloramphenicol O-Acetyltransferase; D013917:Thrombin; D004765:Enteropeptidase