文章摘要
张丽莉,和四梅,王国栋,王艺磊.杂色鲍一种编码CRAD功能域基因的克隆及其在发育和应激中的表达[J].水产学报,2018,42(10):1520~1530
杂色鲍一种编码CRAD功能域基因的克隆及其在发育和应激中的表达
Molecular cloning and expression of a coding CARD gene from Haliotis diversicolor in development and under stresses
投稿时间:2017-09-09  修订日期:2017-10-09
DOI:10.11964/jfc.20170910959
中文关键词: 杂色鲍  Caspase募集功能域(CRAD)  基因表达  发育  应激  RNAi
英文关键词: Haliotis diversicolor  Caspase recruitment domain (CARD)  gene expression  development  stress  RNAi
基金项目:国家自然科学基金(41006105,41176152,3170130721);福建省自然科学基金(2015J01142,2016J01163);福建省高校新世纪优秀人才支持计划(B15138);青岛海洋科学与技术国家实验室开放基金(OF2015NO11)
作者单位E-mail
张丽莉 集美大学水产学院, 农业部东海海水健康养殖重点实验室, 福建 厦门 361021  
和四梅 集美大学水产学院, 农业部东海海水健康养殖重点实验室, 福建 厦门 361021  
王国栋 集美大学水产学院, 农业部东海海水健康养殖重点实验室, 福建 厦门 361021 gdongwang@163.com 
王艺磊 集美大学水产学院, 农业部东海海水健康养殖重点实验室, 福建 厦门 361021 ylwang@jmu.edu.cn 
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中文摘要:
      为探讨Caspase募集功能域(CRAD)在鲍发育和应激中的作用,实验通过比对杂色鲍转录组序列和RACE扩增的方法,获得了杂色鲍一条含CRAD的基因全长cDNA,命名为HdCRAD;采用实时定量PCR的方法获得了该基因在发育和应激中的表达谱。结果显示,HdCRAD的cDNA全长1 371 bp,开放阅读框735 bp,编码244个氨基酸。编码蛋白具有保守的CRAD功能域。该基因在各检测组织中均可表达,在黏液腺中表达量最高;在发育各阶段也均有表达,面盘幼虫晚期表达量最高。在细菌感染、高温或缺氧应激处理后,血细胞的HdCRAD表达水平均有显著性变化。在担轮幼虫时期,RNA干扰该基因会显著提高幼虫畸形率;幼虫的caspase-8和DAD1的表达水平会显著上升,而caspase-3的表达水平会下降。研究表明,该基因参与了鲍幼虫发育过程;在成体免疫、耐高温和抗低氧中发挥了一定作用。
英文摘要:
      In order to analyze the role of Caspase recruitment domain (CARD) of abalone in development and under stress, the full length cDNA of a coding CARD gene was cloned by RACE according to its partial sequence in transcritpome of Haliotis diversicolor. Its expression profile of development and stress was obtained by real time quantitative PCR. The cDNA sequence was named HdCARD and was 1371 bp, with a 735 bp open reading frame encoding a protein of 244 aa (HdCARD) and containing a conserved CARD domain. Quantitative real-time PCR results indicated that HdCARD could be detected in all examined tissues, with the highest expression level in mucus gland. There was also expression of HdCARD in different developmental stages, with highest expression level during late veliger stage. There was a significant difference of HdCARD in hemocytes after a bacterial challenge/thermal/hypoxia stress. The RNAi of HdCARD could increase larval malformation rate in trochophore stage. Moreover, larval caspase-8 and DAD1 significantly rose, but larval caspase-3 significantly reduced. The results suggested that HdCARD might play a role in larval development, immunity, and under thermal stress and hypoxia stress.
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