摘要
为明确中间球海胆己糖激酶的序列信息和表达规律,了解高温-酸化胁迫对其表达和生物活性的影响,以中间球海胆为研究对象,利用cDNA末端快速扩增技术获得中间球海胆已糖激酶基因的全长cDNA序列(SiHK),并利用生物信息学软件分析其序列特征,分析高温-酸化胁迫下中间球海胆肠和性腺组织中SiHK基因的表达及其酶活力变化。结果表明,SiHK基因的cDNA全长2041 bp,编码476个氨基酸,SiHK蛋白的理论等电点为6.44,蛋白质分子质量52.72 kD。生物信息学分析显示,SiHK蛋白氨基酸序列与紫球海胆HK氨基酸序列相似最高。实时荧光定量PCR结果显示,SiHK基因的表达具有组织特异性,其中,肠和性腺组织中SiHK基因的相对表达量较高,而在体腔液和管足中SiHK酶活力较高。高温-酸化胁迫处理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SiHK基因的相对表达量和SiHK酶活性均发生改变。由此表明,高温-酸化胁迫可能通过调控糖代谢关键酶活性对海胆的物质代谢过程产生影响。
In order to clarify the sequence information and expression pattern of hexokinase(HK)in Strongylocentrotus intermedius,and preliminarily understand the effects of high temperature-acidification stress on its expression and activity,Strongylocentrotus intermedius was as material.The full-length cDNA of SiHK in Strongylocentrotus intermedius was cloned and characterized using cDNA ends rapid amplification(RACE)technology and bioinformatics software.And the expression and enzyme activity of SiHK were analyzed in different tissues.The results showed that the total cDNA length of SiHK gene was 2041 bp,encoding 476 amino acids.The theoretical isoelectric point(pI)and molecular weight of SiHK protein were 6.44 and 52.72 kD,respectively.The bioinformatics analysis showed that the amino acid sequence of SiHK protein was the most similar to that of Strongylocentrotus purpuratus HK.The results of qRT-PCR showed that SiHK was expressed in all examined tissues with an obvious tissue specifity,which the relative expression levels in intestine and gonad were higher and the total enzyme activities of SiHK in coelomic fluid and tube feet were higher.Compared with CK,the relative expression of SiHK gene and total enzyme activity of SiHK in intestine and gonad of S.intermedius were changed after high temperature-acidification stress 60 d,which suggested that high temperature-acidification stress might affect metabolism of sea urchins by regulating the expression and enzyme activity of key enzymes of glucose metabolism.
作者
武博琼
崔东遥
焦仁和
宋坚
湛垚垚
常亚青
WU Boqiong;CUI Dongyao;JIAO Renhe;SONG Jian;ZHAN Yaoyao;CHANG Yaqing(Key Laboratory of Mariculture&Stock Enhancement in North China’s Sea,Ministry of Agriculture and Rural Affairs,College of Fisheries and Life Science Dalian Ocean University,Liaoning Dalian 116023,China)
出处
《中国农业科技导报》
CAS
CSCD
北大核心
2022年第7期205-217,共13页
Journal of Agricultural Science and Technology
基金
辽宁省“兴辽英才计划”项目(XLYC2002107)
辽宁省高等学校创新人才支持计划项目(LR2020065)。
关键词
中间球海胆
己糖激酶
基因克隆
高温-酸化胁迫
表达模式
酶活力
Strongylocentrotus intermedius
hexokinase
gene cloning
high temperature-acidification stress
expression pattern
enzyme activity
