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Cloning of SiPFK Gene from Strongylocentrotus intermedius and the Effect of Acidification-High Temperature Stress on Its Expression |
JIAO Ren-He, CUI Dong-Yao, WU Bo-Qiong, SONG Jian, CHANG Ya-Qing, ZHAN Yao-Yao* |
Key Laboratory of Mariculture & Stock Enhancement in North China's Sea, Ministry of Agriculture and Rural Affairs, Dalian Ocean University, Dalian 116023, China |
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Abstract The key enzymes of glycolysis pathway play an important role in regulation and adaptation of echinoderms to the changes of marine environment.In order to clarify the sequence information and expression pattern of phosphofructokinase (PFK) in Strongylocentrotus intermedius, as well as, to understand the effect of "acidification-high temperature" stress on its expression and biological activity, in this study, the rapid-amplification of cDNA ends (RACE) technique was used to clone the full-length cDNA sequence of PFK gene in S. intermedius (SiPFK)(GenBank No. OM780114), and then, the characteristics of SiPFK were analyzed by bioinformatics software, at last, the relative expression pattern of SiPFK and enzyme activity of SiPFK in intestines and gonads of S. intermedius under "acidification-high temperature" stress were investigated. The results showed that, the full- length cDNA of SiPFK gene was 2 787 bp, encoding 840 amino acids, the theoretical isoelectric point of SiPFK protein was 7.48 with a predicted molecular weight of 92.01 kD; The amino acid sequence of SiPFK protein was the most similar to that from S. purpuratus (similarity: 96.20 %); The results of qPCR showed that SiPFK gene specificly expressed in all examined tissue, the relative expression level of SiPFK and total SiPFK enzyme activities in intestines and gonads of S. intermedius were changed after 60 d of "acidification-high temperature" stress, suggesting that "acidification-high temperature" might affect the metabolic process of sea urchins by regulating the expression and activity of key enzymes of glucose metabolism. This study provides an important reference basis for exploring the response of echinoderms to future marine environmental changes.
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Received: 03 December 2021
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Corresponding Authors:
* zhanyaoyao@dlou.edu.cn
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