摘要
为获得罗氏沼虾蜕皮激素受体(ecdysone receptor,EcR)基因编码序列,并检测mRNA表达模式。本实验拟利用克隆测序技术,以肝胰腺为实验素材,扩增EcR基因的编码区;借助生物信息学手段对所得序列进行分析;采用实时荧光定量PCR技术检测EcR基因在肌肉和肝胰腺等8个组织中的m RNA表达量。结果表明,罗氏沼虾EcR基因的编码区长度为1716 bp,共编码571个氨基酸。物种间同源性比较分析发现,罗氏沼虾EcR基因序列与褐虾、黑背陆地蟹、日本沼虾、美洲龙螯虾、大西洋砂招潮蟹、蓝蟹、三疣梭子蟹、中华绒螯蟹和拟穴青蟹的同源性分别为90.7%、84.1%、83%、81%、81%、80.6%、79.8%、77.2%和75.9%。组织表达谱结果显示,EcR基因在所检8种组织中广泛表达,且不同性别相同组织间和相同性别不同组织间EcR的表达均存在不同程度的差异性。其中EcR基因在雄虾的鳃组织中表达量最高,在肌肉组织表达量最低;在雌虾的卵巢组织中高表达,在腹节神经组织中低表达。此外,EcR基因在雌虾的性腺、肠和肝胰腺组织中的表达量极显著或显著高于雄虾(p<0.01或p<0.05),在鳃组织中极显著低于雄虾(p<0.01)。本研究成功克隆了罗氏沼虾的EcR基因,检测到EcR基因在各组织中广泛表达,且在不同性别相同组织和相同性别不同组织中存在显著差异。本实验为进一步研究EcR基因与生长发育调控作用提供了理论依据。
To obtain the coding sequence(CDS)of ecdysone receptor(ECR)gene and detect the expression pattern,cloning and sequencing technology was used to amplify the CDS of EcR gene based on the hepatopancreas of Macrobrachium rosenbergii,bioinformatics tools were adopted to analyze the sequence and the Real-time fluorescence quantitative PCR was applied to detect the expression of EcR gene in eight different tissues.It was shown that open reading frame(ORF)of EcR gene was 1716 bp in length,encoding 571 amino acids.The homology analysis demonstrated that the similarity of EcR with Crangon crangon,Gecarcinus lateralis,Macrobrachium nipponense,Homarus americanus,Leptuca pugilator,Callinectes sapidus,Portunus trituberculatus,Eriocheir sinensis and Scylla paramamosain was 90.7%,84.1%,83%,81%,81%,80.6%,79.8%,77.2%and 75.9%,respectively.The results of expression pattern indicated that EcR was expressed in all tested tissues of Macrobrachium rosenbergii,including abdominal ganglion,stomach,gill,heart,gonad,muscle,hepatopancreas and intestines,and the expression differences were identified in the same tissues of different genders or different tissues of same genders.The highest expression level was observed in gill and ovary,while the lowest in muscle and abdominal ganglion in male and female individuals,respectively.Moreover,the expression in gonad,intestine and hepatopancreas from female prawns were extremely significant higher(p<0.01 or p<0.05),and expression in gill was extremely significant lower than those from male prawns(p<0.01).Taken together,the CDS of EcR gene was successfully cloned,and it was widely distributed in various tissues.Moreover,significant differences were observed in the same tissues between different genders or different tissues between same genders of prawns.The results in the present study will provide a theoretical basis for further research on regulating growth and development of EcR gene in Macrobrachium rosenbergii.
作者
邱庆庆
袁翔
黄光华
蒋钦杨
杨秀荣
姜建萍
蒋和生
Qiu Qingqing;Yuan Xiang;Huang Guanghua;Jiang Qinyang;Yang Xiurong;Jiang Jianping;Jiang Hesheng(College of Animal Science and Technology,Guangxi University,Nanning,530004;Guangxi Institute of Fisheries,Nanning,5300213;Guangxi Botanical Garden of Medicinal Plants,Nanning,530023)
出处
《基因组学与应用生物学》
CAS
CSCD
北大核心
2020年第5期1999-2005,共7页
Genomics and Applied Biology
基金
广西科技重大专项课题(桂科AA17204080-6)
广西自然科学基金项目(2018GXNSFBA281209)共同资助。
关键词
罗氏沼虾
EcR基因
克隆
序列分析
组织表达
Macrobrachium rosenbergii
EcR gene
Cloning
Sequence analysis
Tissue expression
