Molecular cloning and transcriptional analysis of a NPY receptor-like in common Chinese cuttlefish Sepiella japonica  

作　　者：YANG Jingwen XU Yuchao XU Ke PING Hongling SHI Huilai LU Zhenming WU Changwen WANG Tianming 杨静文;徐玉超;许珂;平洪领;史会来;吕振明;吴常文;王天明(National Engineering Research Center for Facilitated Marine Aquaculture,Marine Science College,Zhejiang Ocean University Zhoushan 316022,China;Marine Fisheries Research Institute of Zhejiang Province,Zhoushan 316022,China)

机构地区：[1]National Engineering Research Center for Facilitated Marine Aquaculture,Marine Science College,Zhejiang Ocean University,Zhoushan 316022,China [2]Marine Fisheries Research Institute of Zhejiang Province,Zhoushan 316022,China

出　　处：《Journal of Oceanology and Limnology》2018年第3期892-904,共13页海洋湖沼学报（英文）

基　　金：Supported by the Public Welfare Technical Applied Research Project of Zhejiang Province(No.2017C32074);the International Science&Technology Cooperation Program of China(No.2014DFT30120);the Open Foundation from Marine Sciences in the Most Important Subjects of Zhejiang(No.20130202)

摘　　要：Neuropeptide Y(NPY) has a pivotal role in the regulation of many physiological processes. In this study, the gene encoding a NPY receptor-like from the common Chinese cuttlefish Sepiella japonica( SjNPYR-like) was identified and characterized. The full-length SjNPYR-like cDNA was cloned containing a 492-bp of 5′ untranslated region(UTR), 1 182 bp open reading frame(ORF) encoding a protein of 393 amino acid residues, and 228 bp of 3′ UTR. The putative protein was predicted to have a molecular weight of 45.54 kDa and an isoelectric point(pI) of 8.13. By informatic analyses, SjNPYR-like was identified as belonging to the class A G protein coupled receptor(GPCR) family(the rhodopsin-type). The amino acid sequence contained 12 potential phosphorylation sites and five predicted N-linked glycosylation sites. Multiple sequence alignment and 3D structure modeling were conducted to clarify SjNPYR bioinformatics characteristics. Phylogenetic analysis identifies it as an NPYR with identity of 33% to Lymnaea stagnalis NPFR. Transmembrane properties of SjNPYR-like were demonstrated in vitro using HEK293 cells and the p EGFP-N1 plasmid. Relative quantifi cation of SjNPYR-like mRNA level confi rmed a high level expression and broad distribution of SjNPYR-like in various tissues of female S. japonica. In addition, the transcriptional profile of SjNPYR-like in the brain, liver, and ovary during gonadal development was analyzed. The results provide basic understanding on the molecular characteristics of SjNPYR-like and its potentially physical functions.Neuropeptide Y（NPY） has a pivotal role in the regulation of many physiological processes. In this study, the gene encoding a NPY receptor-like from the common Chinese cuttlefish Sepiella japonica（ SjNPYR-like） was identified and characterized. The full-length SjNPYR-like cDNA was cloned containing a 492-bp of 5′ untranslated region（UTR）, 1 182 bp open reading frame（ORF） encoding a protein of 393 amino acid residues, and 228 bp of 3′ UTR. The putative protein was predicted to have a molecular weight of 45.54 kDa and an isoelectric point（pI） of 8.13. By informatic analyses, SjNPYR-like was identified as belonging to the class A G protein coupled receptor（GPCR） family（the rhodopsin-type）. The amino acid sequence contained 12 potential phosphorylation sites and five predicted N-linked glycosylation sites. Multiple sequence alignment and 3D structure modeling were conducted to clarify SjNPYR bioinformatics characteristics. Phylogenetic analysis identifies it as an NPYR with identity of 33% to Lymnaea stagnalis NPFR. Transmembrane properties of SjNPYR-like were demonstrated in vitro using HEK293 cells and the p EGFP-N1 plasmid. Relative quantifi cation of SjNPYR-like mRNA level confi rmed a high level expression and broad distribution of SjNPYR-like in various tissues of female S. japonica. In addition, the transcriptional profile of SjNPYR-like in the brain, liver, and ovary during gonadal development was analyzed. The results provide basic understanding on the molecular characteristics of SjNPYR-like and its potentially physical functions.

关 键 词：Sepiella japonica NPY receptor-like growth reproduction gene expression 

分 类 号：Q516[生物学—生物化学] Q811.4