杂色鲍caspase-3基因的克隆及其在发育、弧菌感染、高温和缺氧应激中的表达分析  

Cloning and expression analysis of caspase-3 gene in development,immune and stress of variously colored abalone Haliotis diversicolor

作　　者：卢锡琴 张丽莉[1] 黄世玉[1] 王国栋[1] 王艺磊[1] 和四梅[1] LU Xiqin;ZHANG Lili;HUANG Shiyu;WANG Guodong;WANG Yilei;HE Simei(Fisheries College,Key Laboratory of Healthy Mariculture for East China Sea, Ministry of Agriculture and Rural Affairs, Jimei University, Xiamen 361021, China)

机构地区：[1]集美大学水产学院,农业农村部东海海水健康养殖重点实验室,福建厦门361021

出　　处：《大连海洋大学学报》2022年第3期411-419,共9页Journal of Dalian Ocean University

基　　金：国家自然科学基金(31702339)。

摘　　要：为探讨Caspase细胞凋亡通路在杂色鲍Haliotis diversicolor早期发育、免疫及应激过程中的作用,采用cDNA末端快速扩增法克隆出caspase-3基因的全长cDNA序列,并用定量PCR分析了caspase-3基因在各组织、早期发育及经感染副溶血弧菌、高温应激和缺氧应激处理后的表达情况。结果表明:caspase-3基因的cDNA全长为1192 bp,共编码252个氨基酸,该氨基酸序列具有高度保守的P20和P10活性位点;caspase-3基因在杂色鲍成体的各个组织中均有表达,其中血淋巴中的表达量显著高于其他组织(P<0.05);在早期发育阶段中受精卵和2细胞期的表达量最高(P<0.05);经副溶血弧菌感染后,血淋巴中caspase-3的表达量在12 h时显著上调(P<0.05);经高温应激处理后,血淋巴和肝胰腺中caspase-3基因的表达量在24 h时显著上调(P<0.05);经缺氧处理后,鳃组织中caspase-3基因的表达量在96 h时出现显著上调(P<0.05),而血淋巴中的表达量在4 h时显著上调(P<0.05)。研究表明,caspase-3基因在杂色鲍早期发育、免疫和应激中发挥着重要作用。In order to explore the role of caspase apoptosis pathway in early development,immunity and stress of variously colored abalone Haliotis diversicolor,a full-length cDNA sequence of caspase-3 gene was cloned by rapid amplification of cDNA ends.qPCR was used to analyze the expression of caspase-3 gene in various tissues,early developmental stages,infection with Vibrio parahaemolyticus,thermal stress and hypoxia stress.The results showed that the full length of caspase-3 cDNA was 1192 bp,with encoding 252 amino acids.The encoding protein has highly conserved P20 and P10 active sites.There was a constitutive expression in detected tissues and in early developmental stages,with the highest expression in the hemolymph and in the fertilized egg and 2-cell stage(P<0.05),respectively.The expression level of caspase-3 in the hemolymph was significantly increased at 12 h after V.parahaemolyticus infection(P<0.05).After thermal stress,the expression of caspase-3 in hemolymph and hepatopancreas was significantly up-regulated at 24 h(P<0.05).After hypoxia stress,the expression of caspase-3 in gill tissue was significantly up-regulated at 96 h(P<0.05),while the expression of caspase-3 in hemolymph was significantly increased at 24 h(P<0.05).The results suggest that caspase-3 might play an important role in early development,immunity and stress of H.diversicolor.

关 键 词：杂色鲍 CASPASE-3基因 发育 免疫 应激 

分 类 号：S917.4[农业科学—水产科学] Q786[生物学—分子生物学]