Functional identification of the mouse circadian Clock gene by transgenic BAC rescue.
As a complementary approach to positional cloning, we used in vivo complementation with bacterial artificial chromosome (BAC) clones expressed in transgenic mice to identify the circadian Clock gene. A 140 kb BAC transgene completely rescued both the long period and the loss-of-rhythm phenotypes in Clock mutant mice. Analysis with overlapping BAC transgenes demonstrates that a large transcription unit spanning approximately 100,000 base pairs is the Clock gene and encodes a novel basic-helix-loop-helix-PAS domain protein. Overexpression of the Clock transgene can shorten period length beyond the wild-type range, which provides additional evidence that Clock is an integral component of the circadian pacemaking system. Taken together, these results provide a proof of principle that "cloning by rescue" is an efficient and definitive method in mice.
Antoch
M P
MP
National Science Foundation Center for Biological Timing, Department of Neurobiology and Physiology, Northwestern University, Evanston, Illinois 60208, USA.
Song
E J
EJ
Chang
A M
AM
Vitaterna
M H
MH
Zhao
Y
Y
Wilsbacher
L D
LD
Sangoram
A M
AM
King
D P
DP
Pinto
L H
LH
Takahashi
J S
JS
D000818:Animals; D001483:Base Sequence; D056926:CLOCK Proteins; D002874:Chromosome Mapping; D002876:Chromosomes, Bacterial; D002940:Circadian Rhythm / Q000235:genetics* / Q000502:physiology; D003001:Cloning, Molecular; D017931:DNA Primers / Q000235:genetics; D005260:Female; D005816:Genetic Complementation Test; D017403:In Situ Hybridization; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D009154:Mutation; D010641:Phenotype; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D015534:Trans-Activators / Q000235:genetics* / Q000502:physiology
D017931:DNA Primers; D012333:RNA, Messenger; D015534:Trans-Activators; D056926:CLOCK Proteins; C538944:Clock protein, mouse