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Analysis of the circadian clock gene period in the sheep blow fly Lucilia cuprina

Published online by Cambridge University Press:  01 June 2000

G. R. WARMAN
Affiliation:
Present address: School of Biological Sciences, University of Surrey, Guildford GU2 5XH, U.K.
R. D. NEWCOMB
Affiliation:
HortResearch, Mt Albert Research Centre, Private Bag 92169, Auckland, New Zealand
R. D. LEWIS
Affiliation:
School of Biological Sciences, University of Auckland, Private Bag 92019, Auckland, New Zealand
C. W. EVANS
Affiliation:
School of Biological Sciences, University of Auckland, Private Bag 92019, Auckland, New Zealand

Abstract

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We have isolated a homologue of the period (per) gene from the Australian sheep blow fly, Lucilia cuprina, as part of a comparative approach to the analysis of dipteran circadian systems. Sequence analysis of the 4 kb per cDNA revealed the conservation of three functional domains, namely the PAS dimerization motif, and the nuclear and cytoplasmic localization domains. A fourth domain, the threonine–glycine (TG) repeat region, is also conserved in L. cuprina per but has been severely truncated. No length variation was found in the TG repeat of L. cuprina or L. sericata collected from several different latitudinal zones. Expression analysis indicated a diel oscillation in per mRNA in LD 12[ratio ]12 with a period of 24 h and a peak at Zt 12. PER-immunoreactive protein oscillations were also demonstrated, with peak immunoreactivity lagging approximately 3 h behind peak mRNA levels. These results show the existence of a Drosophila-like circadian system in a calliphorid fly. They also provide evidence for the conservation of per function across the Diptera, and confirm the relevance of the Drosophila system as a model for fly circadian rhythms.

Type
Research Article
Copyright
© 2000 Cambridge University Press