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| Cloning and Expression Response of PcCyc Gene to Photoperiod in Red Swamp Crayfish Procambarus clarkii |
| XIE Wei1, JIANG Qichen2, SUN Bin1, YANG Ying1, LI Peng1, YAN Jie1, ZHOU Kaiya1 |
1.Jiangsu Key Laboratory for Biodiversity & Biotechnology, College of Life Sciences, Nanjing Normal University, Nanjing 210023, China;
2.Jiangsu Province Fisheries Research Institute, Nanjing 210017, China |
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Abstract In this study, a partial cDNA sequence of cycle gene (PcCyc), one of the core genes of the circadian clock, was cloned from red swamp crayfish Procambarus clarkii with body length of 9—11 cm by rapid amplification of cDNA ends (RACE) technology, and then the mRNA expression levels of PcCyc gene were detected in brain, eye stalk and hepatopancreas of the red swamp crayfish exposed to various photoperiods of 12L∶12D, 0L∶24D, and 0L∶72D under full-spectrum fluorescent lamps at 7:00, 11:00, 15:00, 19:00, 23:00, 03:00 and 7:00 the next day by real-time quantitative PCR. The results showed that the mRNA of PcCyc gene had a complete open reading frame (ORF) of 2010 base pairs encoding 669 amino acids, and the amino acid sequence of PcCYC had the maximal similarity (94%) to that of the red claw crayfish Cherax quadricarinatus, with the protein molecular weight of about 73.963 ku, the theoretical isoelectric point of 6.56, which is a hydrophobic non-secreted protein with no signal peptide. The structural domain analysis revealed that there were a BHLH/PAS domain, including one HLH domain and two PAS domains in the PcCYC protein, with secondary structure of 11 alpha-helices, 12 beta-sheets, and 24 random coils. The PcCyc gene was shown to be 24-hour rhythm fluctuation in both eyestalk and hepatopancreas, no matter what photoperiod was. However, the rhythmic fluctuation was broken in the brain as the photoperiod changed. The findings provide the foundation for further exploration of the role and mechanism of the Cyc gene in circadian clock of the red swamp crayfish.
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Received: 27 September 2019
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