摘要
利用RACE方法获得了脊尾白虾(Exopalaemon carinicauda)酚氧化酶原(Prophenoloxidase,proPO)基因。该基因cDNA全长3092bp,开放阅读框2013bp,编码671个氨基酸,其预测分子量为74.82kDa。脊尾白虾酚氧化酶原基因推导的氨基酸序列与其他虾类酚氧化酶原氨基酸序列同源性较高(65%—78%),该氨基酸序列具有铜离子结合位点等酚氧化酶原基因所具有的典型特征位点。组织表达分析结果表明,proPO基因在脊尾白虾血细胞中表达量最高,其次是鳃和肝胰腺组织,在肌肉中几乎不表达。利用荧光定量PCR分析了不同盐度胁迫下脊尾白虾血细胞和鳃组织中的表达变化规律,结果发现在不同盐度胁迫后血淋巴proPO表达量显著高于对照组,盐度胁迫初期鳃组织proPO表达量显著高于对照组。本研究结果表明脊尾白虾酚氧化酶原基因参与了盐度胁迫引起的机体应激反应。
The cDNA sequence of prophenoloxidase(proPO) gene was cloned from Exopalaemon carinicauda using RT-PCR and RACE method. The full-length cDNA of proPO consisted of 3092 bp with a 2013 bp open reading frame(ORF), which encoded 671 amino acids. The predicted molecular mass of proPO protein is 74.82 kDa. The deduced amino acid sequence shows higher homology(65%—78%) to other shrimps with characteristic proPO. Tissue specific expression analysis showed that the expression level of proPO mRNA is highest in haemocytes and lowest in muscle. Real time-PCR analysis showed that the expression level of proPO gene is significantly higher in haemocytes than that of the control after 24 h exposure. The expression level of proPO mRNA in gills is significantly higher than that of the control at the early stage of salinity stress. The results show that the proPO gene might be involved in the stress response in E. carinicauda.
出处
《海洋与湖沼》
CAS
CSCD
北大核心
2014年第2期299-306,共8页
Oceanologia Et Limnologia Sinica
基金
国家高技术研究发展计划(863)项目资助
"主要养殖甲壳类良种培育"
2012AA10A409号
国家虾产业技术体系项目资助
CARS-47号
公益性行业(农业)科研专项课题资助
201103034号
中国水产科学研究院基本科研业务费项目资助
2013A0701号
关键词
脊尾白虾
酚氧化酶原
基因克隆
表达
盐度胁迫
Exopalamon carincauda
prophenoloxidase
cloning
expression
salinity stress
