刺参甜菜碱高半胱氨酸甲基转移酶1基因(bhmt1)的克隆、分析及其在低盐胁迫下的表达变化  

Cloning of Betaine Homocysteine Methyltransferase 1 (bhmt1) Gene from Apostichopus japonicus and Its Expression Pattern under Low Salinity Stress

作　　者：赵业 卜少阳 王涵 崔皓 郝志军 许润润 李诗浩 程亚秋 马恬 ZHAO Ye;BU Shao-yang;WANG Han;CUI hao;HAO Zhi-jun;XU Run-run;LI Shi-hao;CHENG Ya-qiu;MA Tian(School of Ocean,Yantai University,Yantai 264005,China;Technology Center,Yantai Customs,Yantai 265699,China)

机构地区：[1]烟台大学海洋学院,山东烟台264005 [2]烟台海关技术中心,山东烟台265699

出　　处：《烟台大学学报（自然科学与工程版）》2022年第4期434-442,共9页Journal of Yantai University(Natural Science and Engineering Edition)

基　　金：国家自然科学基金资助项目(41906098);烟台大学大学生创新创业训练计划项目。

摘　　要：研究克隆获取了刺参(Apostichopus japonicus)甜菜碱高半胱氨酸甲基转移酶bhmt1(betaine homocysteine methyltransferase 1)基因全长序列,并研究了该基因在不同低盐胁迫下的时空表达模式。结果表明:刺参bhmt1基因的全长cDNA为2328 bp,开放阅读框为1251 bp,共编码416个氨基酸,包括BHMT的典型基序结构GGCCGFEPYHI及三个可能作为底物结合位点的锌指结构序列。系统进化树分析表明刺参BHMT1与已报道物种相似度为66.08%~66.67%,且进化地位与刺参生物学分类地位基本一致。荧光实时定量分析表明bhmt1基因在刺参多种组织均有表达,且在呼吸树中表达最高。低盐胁迫实验表明bhmt1基因在不同低盐胁迫条件中刺参肠道和呼吸树组织中出现了显著上调表达(P<0.05),表明其可能在适应环境渗透压变化中起重要作用。In this study, the complete cDNA sequence of the betaine homocysteine methyltransferase 1(bhmt1) gene is cloned from the sea cucumber Apostichopus japonicus, and its temporal and spatial expression pattern is further detected and analyzed under different low salinity stress. The results show that the full-length cDNA of the bhmt1 gene is 2328 bp, with an open reading frame of 1251 bp encoding 416 amino acids, including the typical motif structure of BHMT, GGCCGFEPYHI, and three zinc finger structure sequences which may be used as substrate binding sites. The phylogenetic tree analysis suggests that the evolutionary status of bhmt1 in A. japonicus is basically consistent with the biological taxonomic status of sea cucumber. RT-qPCR analysis shows that bhmt1 gene expression is found in different tissues of A. japonicus, and is the highest in arbor respiratoris. The expression of bhmt1 gene is found to be up-regulated in intestine and respiratory tree tissues under different low salinity stress, suggesting its important role in the adaption to ambient osmotic pressure.

关 键 词：刺参 bhmt1基因 基因克隆 实时定量PCR分析 低盐胁迫 

分 类 号：S917.4[农业科学—水产科学]