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中华蛸章鱼胺受体OARβ2R基因克隆及表达分析
肖懿哲1, 陈小灵2, 孙玉龙3, 张子平3, 王艺磊2, 朱友芳1
1.莆田市水产科学研究所;2.集美大学水产学院;3.福建农林大学动物科学学院
摘要:
为了解中华蛸(Octopus sinensis)受精卵的孵化及幼体发育机制,本研究对其章鱼胺受体基因(OsOARβ2R)进行克隆和生物信息学分析,同时研究了OsOARβ2R在不同组织/器官、刚孵岀的幼体不同饥饿时间及不同的胚胎发育时期OsOARβ2R表达水平的变化。结果为:OsOARβ2R开放阅读框全长为1158 bp,编码385个氨基酸,有7个跨膜结构域,具有G蛋白偶联受体的共性,其中TM3、TM5、TM6、TM7四个跨膜结构相对保守。经氨基酸同源对比及构建系统进化树分析,其与加州双斑蛸(O. bimaculoides)的OAR的一致性最高。荧光定量PCR结果显示,在成体10个组织/器官中,OsOARβ2R在后唾液腺中表达水平最高,其次是脑和小肠;在幼体饥饿实验中,随着饥饿时间的推移,OsOARβ2R的表达水平在饥饿2d后显著降低,紧接着在饥饿3d时表达水平显著升高,且表达水平达到最高,之后表达水平开始回落;OsOARβ2R在中华蛸整个胚胎发育周期均可检测到,且在多细胞期表达水平最高,在初孵幼体表达水平最低。这些结果为研究中华蛸受精卵的孵化及幼体发育机制,提高其人工育苗的效率提供了基础资料。
关键词:  中华蛸  章鱼胺受体  OsOARβ2R  饥饿  胚胎发育
DOI:
分类号:Q 785; S 968.3????????????????????????????
基金项目:中央引导地方科技发展专项(020L3011); 福建省科技厅项目(2021S21010093); 莆田市科技局项目(2020NJJ005)。
Cloning and expression analysis of OARβ2R gene from Octopus Sinensis
Xiao Yi-zhe1, Chen Xiao-ling2, Sun Yu-long3, Zhang Zi-ping3, Wang Yi-lei2, Zhuyoufang1
1.Putian Municipal Institute of Fishery Science;2.Fisheries College, Jimei University;3.College of Animal Science, Fujian Agriculture and Forestry University
Abstract:
In order to understand the mechanism of hatching and larval development of Octopus sinensis, its OsOARβ2R gene was cloned, and bioinformatics analysis of the gene was carried out. Meanwhile, the expression of OsOARβ2R in different tissues/organs, at different starvation periods of newly hatched larvae, and different embryonic development stages were analyzed. The results are: the open reading frame of OsOARβ2R is 1158 bp, encoding 385 amino acids and containing 7 transmembrane domains (TMs), which have the commonness of G protein-coupled receptors. Among them, the TM3, TM5, TM6 and TM7 are relatively conservative. OsOARβ2R has the highest consistency with the OAR of O. bimaculoides through amino acid homology comparison and phylogenetic tree analysis. The results of quantitative PCR showed that the expression of OsOARβ2R was expressed widely in 10 examined tissues/organs with the highest expression level in posterior salivary gland, followed by brain and intestine; In the larval starvation experiment, the expression of OsOARβ2R decreased significantly after 2 days of starvation, then increased significantly and reached the highest after 3 days of starvation, and then decline to normal level; In the embryonic development, the expression level of OsOARβ2R was the highest in multicellular stage and the lowest in newly hatched larvae. These results provide basic data for studying the hatching and larval development mechanism of fertilized eggs of O. sinensis, and improving the efficiency of artificial seedling for it.
Key words:  Octopus Sinensis, OARβ2R, starvation, embryonic development
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