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2025年4月6日 星期日
农业生物技术学报  2022, Vol. 30 Issue (10): 1962-1975    DOI: 10.3969/j.issn.1674-7968.2022.10.011
  研究论文与报告 本期目录 | 过刊浏览 | 高级检索 |
中间球海胆SiPFK基因的克隆及酸化-高温胁迫对其表达的影响
焦仁和, 崔东遥, 武博琼, 宋坚, 常亚青, 湛垚垚*
大连海洋大学 农业农村部北方海水增养殖重点实验室,大连 116023
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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摘要 糖酵解途径中的关键酶磷酸果糖激酶(phosphofructokinase, PFK)在棘皮动物调节和适应海洋环境变化中发挥重要作用。为明确中间球海胆(Strongylocentrotus intermedius) PFK基因的序列信息和表达规律,初步了解“酸化-高温”胁迫对其表达和生物活力的影响,本研究通过cDNA末端快速扩增(rapid-amplification of cDNA ends, RACE)技术获得中间球海胆磷酸果糖激酶基因SiPFK的全长cDNA序列(GenBank No. OM780114),并利用生物信息学软件分析其序列特征,在此基础上,比较分析了“酸化-高温”胁迫条件下,中间球海胆的肠和性腺组织中SiPFK基因的相对表达及其酶活力变化情况。结果表明,SiPFK基因的cDNA全长为2 787 bp,编码840个氨基酸,SiPFK蛋白理论等电点为7.48,蛋白质分子质量为92.01 kD;生物信息学分析显示,SiPFK蛋白氨基酸序列与紫球海胆(S. purpuratus) PFK氨基酸序列最为相似,相似度为96.20%;qPCR和酶活力测定结果显示,SiPFK基因的相对表达和总酶活力的组织特异性较为明显,“酸化-高温”胁迫60 d后,与对照组相比,处理组中间球海胆的肠和性腺组织中SiPFK基因的相对表达量和SiPFK酶活力均发生了改变,推测“酸化-高温”胁迫可能通过调控糖代谢关键酶活力对海胆的物质代谢过程产生影响。本研究为探究棘皮动物响应未来海洋环境变化提供重要参考依据。
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焦仁和
崔东遥
武博琼
宋坚
常亚青
湛垚垚
关键词 中间球海胆磷酸果糖激酶(PFK)基因克隆"酸化-高温"胁迫表达模式酶活力    
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.
Key wordsStrongylocentrotus intermedius    Phosphofructokinase (PFK)    Gene cloning    "Acidification-high temperature" stress    Expression pattern    Enzyme activity
收稿日期: 2021-12-03     
ZTFLH:  S968.9  
  Q786  
基金资助:辽宁省高等学校创新人才支持计划(LR2020065); 辽宁省“兴辽英才计划” (XLYC2002107)
通讯作者: *zhanyaoyao@dlou.edu.cn   
引用本文:   
焦仁和, 崔东遥, 武博琼, 宋坚, 常亚青, 湛垚垚. 中间球海胆SiPFK基因的克隆及酸化-高温胁迫对其表达的影响[J]. 农业生物技术学报, 2022, 30(10): 1962-1975.
JIAO Ren-He, CUI Dong-Yao, WU Bo-Qiong, SONG Jian, CHANG Ya-Qing, ZHAN Yao-Yao. Cloning of SiPFK Gene from Strongylocentrotus intermedius and the Effect of Acidification-High Temperature Stress on Its Expression. 农业生物技术学报, 2022, 30(10): 1962-1975.
链接本文:  
http://journal05.magtech.org.cn/Jwk_ny/CN/10.3969/j.issn.1674-7968.2022.10.011     或     http://journal05.magtech.org.cn/Jwk_ny/CN/Y2022/V30/I10/1962
 
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