摘要
磷脂肌醇3-激酶(phospoinositide-3 kinases,PI3Ks)是真核生物中的一类关键调控因子,通过影响细胞存活、细胞迁移、细胞增殖等进而调控机体生长代谢,但迄今在日本沼虾中未见报道。本研究克隆了日本沼虾(Macrobrachium nipponense)PI3K基因mRNA全长序列,并对其表达进行了初步分析。结果表明:日本沼虾PI3K基因mRNA全长3284 bp,其中5’UTR为138 bp,3’UTR为365 bp,开放阅读框(open reading frame,ORF)长2778 bp,编码926个氨基酸。结构域分析表明其含有PI3K家族第Ⅳ类基因典型结构域PI3K C2、PI3Ka、PI3Kc;故将其命名为PI3Kc3。氨基酸序列同源性分析表明日本沼虾的PI3Kc3基因与凡纳滨对虾(Penaeus vannamei)的同源性最高,达71.27%。半定量PCR和荧光定量PCR结果显示,PI3Kc3基因在日本沼虾的眼睛、大脑、肝胰腺等组织以及受精卵至成虾各个发育时期均有表达,且表达量间无显著差异(P>0.05);同时,检测规格差异显著的全同胞雄虾个体第二步足、肌肉和肝胰腺组织表明大小规格间各组织表达无显著差异(P>0.05)。本研究初步分析了日本沼虾PI3Kc3基因结构及其表达分布,为进一步探讨日本沼虾生长调控提供分子基础。
Phospoinositide-3 kinases(PI3Ks)are a class of key regulators in eukaryotes.They regulate the growth and metabolism of the body by affecting cell survival,cell migration and cell proliferation,but so far no reports have been reported in Macrobrachium nipponense.In this study,the full length sequence of PI3K gene mRNA of M.nipponense was cloned,and its expression was analyzed.The results showed that the total length of PI3K gene mRNA of M.nipponense was 3284 bp,of which 5’UTR was 138 bp and 3’UTR was 365 bp.The open reading frame(ORF)was 2778 bp and encoded 926 amino acids.Domain analysis showed that it contained PI3K family classⅣgene typical domainsPI3K C2,PI3Ka,PI3Kc.Therefore,it was named PI3Kc3.The homology analysis of amino acid sequence showed that the PI3Kc3 gene of M.nipponense had the highest homology with Penaeus vannamei(71.27%).The results of semi-quantitative PCR and fluorescent quantitative PCR showed that the PI3Kc3 gene was expressed in the eyes,brain,hepatopancreas and other tissues of the M.nipponense,and it was also expressed at all stages of development from fertilized eggs to adult shrimp,which there was no significant difference in their expression levels(P>0.05).The second pereiopod,muscle and hepatopancreas tissues of full-sibling male with significant differences in size were detected,and the expression was no significant difference between the tissues of different sizes(P>0.05)This study preliminarily analyzed the structure and expression distribution ofPI3Kc3 gene of M.nipponense,providing a molecular basis for further discussion on the growth regulation of M.nipponense.
作者
朱永明
武源清
刘志伟
黄旭雄
Zhu Yongming;Wu Yuanqing;Liu Zhiwei;Huang Xuxiong(National Demonstration Center for Experimental Fisheries Science Education,Shanghai Ocean University,Shanghai,201306;Shanghai Engineering Research Center of Aquaculture,Shanghai Ocean University,Shanghai,201306)
出处
《基因组学与应用生物学》
CAS
CSCD
北大核心
2021年第9期2916-2924,共9页
Genomics and Applied Biology
基金
国家自然科学基金项目(31101050)
上海市科学技术委员会项目(14320502000)
上海市教委创新计划(13YZ094)共同资助
关键词
日本沼虾
生长调控
PI3K基因
同源性
表达分析
Macrobrachium nipponense
Growth regulation
PI3K gene
Homology
Expression analysis
