Cloning and expression of mouse-brain calmodulin as an activator of Bordetella pertussis adenylate cyclase in Escherichia coli.
Cloning of higher eukaryotic genes has seldom been performed by complementation of a defective prokaryotic function. This is especially true in the case of functions that are normally absent from the prokaryotic host. We demonstrate here that it is possible to identify by complementation the cDNA from mouse brain, which encodes calmodulin (CaM) synthesis, in spite of the fact that the recipient strain, Escherichia coli, does not normally harbour a CaM function. A three-component cloning procedure in which a gene product requiring CaM for activity, adenylate cyclase from the pathogen Bordetella pertussis, was used to screen a cDNA library for cAMP synthesis in E. coli. The nucleotide sequence of the corresponding cDNA is also reported.
Danchin
A
A
Régulation de l'Expression Génétique, Institut Pasteur, Paris, France.
Sezer
O
O
Glaser
P
P
Chalon
P
P
Caput
D
D
D000262:Adenylyl Cyclases / Q000096:biosynthesis* / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001886:Bordetella pertussis / Q000201:enzymology* / Q000235:genetics; D001921:Brain / Q000378:metabolism; D002147:Calmodulin / Q000096:biosynthesis / Q000235:genetics* / Q000502:physiology; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000235:genetics; D004789:Enzyme Activation; D004926:Escherichia coli / Q000235:genetics; D005798:Genes, Bacterial; D051379:Mice; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000235:genetics
D002147:Calmodulin; D004269:DNA, Bacterial; D012333:RNA, Messenger; D000262:Adenylyl Cyclases