CgLOX4基因在长牡蛎幼虫发育中的表达特征  

Expression characteristics of CgLOX4 gene in larval development of Pacific oyster(Crassostrea gigas)

作　　者：胡悦 冷金源 孙洁洁 王玲玲 宋林生 HU Yue;LENG Jinyuan;SUN Jiejie;WANG Linging;SONG Linsheng(Dalian Key Laboratory of Aquatic Animal Diseases Prevention and Control,Liaoning Key Laboratory of Marine Animal Immunology and Epidemic Pre-vention and Control,Dalian Ocean University,Dalian 116023,China)

机构地区：[1]大连海洋大学,辽宁省海洋动物免疫学与疫病防控重点实验室,大连市水产养殖动物病害防控重点实验室,辽宁大连116023

出　　处：《大连海洋大学学报》2024年第1期38-47,共10页Journal of Dalian Ocean University

基　　金：国家自然科学基金(32222086,32173002);大连市优秀青年科技人才项目(2022RY01)。

摘　　要：为研究长牡蛎(Crassostrea gigas)LOX4基因(CgLOX4)在幼虫发育中的表达特征,采用PCR技术扩增CgLOX4的全长cDNA序列,利用荧光定量PCR和整体免疫荧光技术检测CgLOX4在幼虫不同发育时期的表达特征。结果表明:CgLOX4的cDNA全长为1 862 bp,开放阅读框为834 bp,可编码277个氨基酸,等电点为8.36,预测其氨基酸序列只含有一个保守的HOX结构域;CgLOX4在长牡蛎成体各组织中均有表达,其中在闭壳肌中的表达量显著高于肝胰腺、鳃、血淋巴、性腺和唇瓣组织(P<0.05);CgLOX4在所检测的早期胚胎中均有表达,其中在受精卵和4细胞期的表达量相对较高;CgLOX4阳性信号主要分布在担轮幼虫壳形成区、D型幼虫内脏团和壳顶幼虫消化腺中。研究表明,CgLOX4是长牡蛎早期发育过程中较早激活的转录因子之一,可能在调控幼虫壳形成和消化器官形成等方面发挥重要作用。To analyze the expression characteristics of CgLOX4 during larval development,the full-length cDNA se-quence of LOX4 was amplified from Pacific oyster(Crassostrea gigas)by using PCR,and the expression character-istics of CgLOX4 in different stages of larval development were further analyzed by qRT-PCR and whole-mount im-munofluorescence experiments.It was found that the CgLOX4 gene had full-length cDNA sequence of 1862 bp with an open reading frame of 834 bp encoding 277 amino acids,with the isoelectric point of 8.36.The amino acid se-quence of CgLOX4 gene contained a conserved HOX domain,and CgLOX4 gene was expressed in all the tested tis-sues of adult oyster,with significantly higher expression level in the adductor muscle than that in hepatopancreas,gills,hemolymph,gonad and labial palps tissue(P<0.05).CgLOX4 was expressed in all the examined early em-bryos,with relatively high expression level in fertilized eggs,and four-cells.The whole-mount immunofluorescence assay revealed that the positive signals of LOX4 were mainly distributed in the shell field of trochophore,the viscer-al mass of D-shape larvae and the digestive gland of umbo larvae.The findings indicate that CgLOX4 gene is one of the transcription factors activated earlier in the early development process of the oyster,and may play an important role in regulating the formation of larval shell and digestive organ.

关 键 词：长牡蛎 LOX4 幼虫发育 分布特征 

分 类 号：S917.4[农业科学—水产科学]