Screening for Mutants in the Output Pathway of the Circadian Clock in Chlamydomonas Reinhardtii
The circadian clock is a basic component of biological systems and has been found in many organisms. It is composed by three major components: the input pathway, the oscillator and the output pathway. The purpose of this research is to study the output pathway of the circadian clock in Chlamydomonas reinhardtii strain Carnil by screening mutants, which were generated by insertional mutagenesis via glass bead transformation. The plasmid pSP124S containing the ble marker was used to introduce mutations. The CABII-1 gene has been reported to show a circadian rhythm in expression. The reporter gene ARS2 that was transcriptionally fused to the promoter of the CABII-1 gene was then analyzed. The screening process was optimized to be less time-consuming. Eighteen mutants with significantly low ARS2 accumulated expressions were found from 1004 transformants, which implies that there may be an activator involved in the circadian phenotype of CABII-1.
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