| 摘要: |
| 为研究脊尾白虾蜕皮抑制激素基因(MIH)在环境适应中的作用, 本文采用RACE技术从蜕皮间期脊尾白虾眼柄中克隆获得MIH-like基因全长cDNA序列, 命名为EcMIH。该基因全长1218bp, 包括360bp的开放阅读框, 420bp的5'端非编码区和394bp的3'端非编码区;开放阅读框编码120个氨基酸, 包括41个氨基酸组成的信号肽和79个氨基酸组成的成熟肽, 预测蛋白分子量为13.71kDa, 理论等电点pI为8.02。同源性比较分析发现, 脊尾白虾MIH-like基因与日本沼虾和罗氏沼虾同源性最高, 分别为87%和86%。荧光定量PCR分析结果表明, 脊尾白虾MIH-like基因在眼柄中的表达量最高, 在卵巢中的表达量最少, 而在肝脏、肌肉、鳃和血细胞中不表达。环境胁迫后该基因具有明显的时序性。pH6.5胁迫后24h和48h, 眼柄和腹神经节中MIH-like基因的表达量较对照组均显著增加(P<0.05);4mg/L氨氮浓度组胁迫后12h和24h, 眼柄和腹神经节中MIH-like基因的表达量较对照组均显著增加(P<0.05);盐度39胁迫后12h和24h, 眼柄和腹神经节中MIH-like基因的表达量较对照组均显著增加(P<0.05)。本研究结果表明脊尾白虾mih基因参与了环境胁迫后的机体应激反应。 |
| 关键词: 脊尾白虾 蜕皮抑制激素 基因表达 环境胁迫 |
| DOI：10.11693/hyhz20141100332 |
| 分类号: |
| 基金项目:国家高技术研究发展计划(863项目),2012AA10A409号;国家虾产业技术体系-北方养殖岗位项目,CARS-47号;国家自然科学基金项目,31472275号;公益性(农业)行业专项项目,201103034号。 |
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| CLONING AND EXPRESSION OF MOLT-INHIBITING HORMONE GENE FROM EXOPALAEMON CARINICAUDA UNDER ENVIRONMENTAL STRESSES |
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ZHANG Mei1,2, LIU Ping2, LI Ji-Tao2, LI Jian2
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1.Ocean University of Dalian, Dalian 116023, China;2.Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China
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| Abstract: |
| We cloned the full-length MIH (molt-inhibiting hormone)-like cDNA from the eyestalk ganglia of ridgetail white prawn Exopalaemon carinicauda, and named as EcMIH by rapid amplification of cDNA ends (RACE).The full-length of MIH cDNA was 1218bp containing a 420bp 5' untranslated region (5'UTR), a 438 bp 3' untranslated region (3'UTR) and a 360bp open reading frame (ORF) encoding 120 amino acids consisting of a 41-residue signal peptide and a 79-residue mature peptide;the molecular mass was 13.71 kDa and the pI (isoelectric point) was 8.02.Sequence analysis indicated that the EcMIH was in homology with those of Marsupenaeus japonicas and Macrobrachium rosenbergii, in 87% and 86% identity, respectively.Tissue expression analysis showed that the EcMIH mRNA expressed at the highest level in eyestalk while extremely low in ovary, and not expressed in liver, muscle, gill, and blood cells.Under different environmental stresses, the MIH-like gene expression level in eyestalk and ventral ganglion significantly increased (P<0.05) compared to the control (i.e., in pH 6.5 group for 24h and 48h, in 4mg/L NH4Cl group for 12h and 24h, and in salinity 39 group for 12h and 24h).Therefore, the EcMIH is involved in response against environmental stresses. |
| Key words: Exopalaemon carinicauda molt-inhibiting hormone (MIH) gene expression environmental stress |
