真蛸FAXDC2基因的克隆及其表达分析  

Cloning and expression of FAXDC2 in Octopus vulgaris

作　　者：肖懿哲 姚成杰 朱友芳 孙玉龙 张子平[3] 王艺磊[2] XIAO Yi-zhe;YAO Cheng-jie;ZHU You-fang;SUN Yu-long;ZHANG Zi-ping;WANG Yi-lei(Putian Municipal Institute of Fishery Science,Putian 351100,China;Fisheries College,Jimei University,Xiamen 361021,China;College of Animal Science,Fujian Agriculture and Forestry University,Fuzhou 350002,China)

机构地区：[1]莆田市水产科学研究所,福建莆田351100 [2]集美大学水产学院,福建厦门361021 [3]福建农林大学动物科学学院,福建福州350002

出　　处：《海洋科学》2019年第8期56-63,共8页Marine Sciences

基　　金：国家自然科学基金资助项目(41676161,31672681)~~

摘　　要：FAXDC2(fatty acid hydroxylase domain-containing protein 2)是脂肪酸羟化酶家族中的成员,在脂肪酸的合成与代谢中发挥重要功能。真蛸(Octopus vulgaris)是中国南方近海的经济种类,为了研究饥饿对真蛸FAXDC2(OvFAXDC2)基因表达的影响,本研究从课题组真蛸转录组数据库中筛选获得OvFAXDC2基因序列。采用从头至趾方法进行验证,最终获得OvFAXDC2的cDNA全长序列共1384 bp。它包含100 bp的5′非编码区(UTR)、228 bp的3′UTR和1056 bp的开放阅读框(ORF),ORF共编码351个氨基酸,预测其分子量为3.98 kDa,理论等电点为8.93。系统进化树分析显示,真蛸和加州双斑蛸(O.bimaculoides)聚为一支,而与其他无脊椎动物分开。实时荧光定量PCR分析表明,OvFAXDC2在消化腺中表达量最高,在鳃和鳃心中次之。在幼体饥饿试验中,随着时间的推移,OvFAXDC2表达量先升后降,在第三天达到最高,表明OvFAXDC2表达量的变化趋势可作为幼体代谢是否正常的一个指标。Fatty acid hydroxylase domain-containing protein 2(FAXDC2)belongs to the fatty acid hydroxylase super-family and plays vital roles in fatty acidsynthesis and metabolism.Octopus vulgaris is an economically important spe-cies found in the coastal waters of southern China.This study investigated the effects of starvation on OvFAXDC2 ex-pression in O.vulgaris using the cDNA sequence(1384 bp)of OvFAXDC2 obtained from the transcriptome database of O.vulgaris and confirmed by PCR in a head-to-toe manner.This sequence contains a 100-bp 5′untranslated region(UTR),228-bp 3′UTR,and 1056-bp open reading frame encoding 351 amino acids.The molecular weight of the protein is 3.98 kDa,and its theoretical isoelectric point is 8.93.Phylogenetic tree analysis revealed that O.vulgaris and O.bimaculoides were clustered and separated from other invertebrates.Real-time fluorescence quantitative PCR revealed that OvFAXDC2 showed the highest expression in the digestive gland,followed by the gill and gill heart.During lar-valstarvation,OvFAXDC2 expression increased initially and then decreased,reaching the highest level on the third day.Results of this study suggest OvFAXDC2 expression patterns can be used as an indicator of normal larval metabolism.

关 键 词：真蛸(Octopus vulgaris) FAXDC2基因 克隆 组织表达 饥饿 实时荧光定量PCR 

分 类 号：Q785[生物学—分子生物学] S968.3[农业科学—水产养殖]