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黎军胜, 李建林, 吴婷婷. 罗非鱼胰蛋白酶ⅠmRNA克隆与分析[J]. 水生生物学报, 2006, 30(2): 209-213.
引用本文: 黎军胜, 李建林, 吴婷婷. 罗非鱼胰蛋白酶ⅠmRNA克隆与分析[J]. 水生生物学报, 2006, 30(2): 209-213.
LI Jun-Sheng, LI Jian-Lin, WU Ting-Ting. CLONING AND SEQUENCING OF TRYPSINⅠMRNA IN TILAPIA[J]. ACTA HYDROBIOLOGICA SINICA, 2006, 30(2): 209-213.
Citation: LI Jun-Sheng, LI Jian-Lin, WU Ting-Ting. CLONING AND SEQUENCING OF TRYPSINⅠMRNA IN TILAPIA[J]. ACTA HYDROBIOLOGICA SINICA, 2006, 30(2): 209-213.

罗非鱼胰蛋白酶ⅠmRNA克隆与分析

CLONING AND SEQUENCING OF TRYPSINⅠMRNA IN TILAPIA

  • 摘要: 应用3′/5-′RACE方法对尼罗罗非鱼(Oreochromis niloticus)和奥利亚(Oreochromis aureus)罗非鱼胰蛋白酶ⅠmR-NA进行了克隆和测序,序列分析表明,尼罗罗非鱼和奥利亚罗非鱼胰蛋白酶ⅠmRNA序列全长分别为863bp和858bp,序列中均含有738个核苷酸组成的开放阅读框,编码长度为245个氨基酸残基的胰蛋白酶Ⅰ。胰蛋白酶Ⅰ氨基末端均存在信号肽和激活肽序列,序列中均具有与催化活性必须的高度保守的催化三联体氨基酸残基(His、Asp、Ser)和构成二硫键的12个半胱氨酸残基,以及决定底物特异性的保守性氨基酸残基即天冬氨酸残基和S1结合袋。南极鱼(Patanotothenia magellanica)、大西洋鲑(Salmo salar)、日本鲽(Paralichthys olivaceus)、大西洋鳕(Gadusmorhua)、牛和人类胰蛋白酶mRNA序列以及氨基酸序列与奥利亚罗非鱼相比较同源性分别为58.3%—72.5%和63.3%—76.1%,与尼罗罗非鱼相比较同源性分别为59.1%—73.1%和63.6%—75.6%。奥利亚罗非鱼和尼罗罗非鱼胰蛋白酶mRNA序列以及氨基酸序列同源性分别为96.2%和99.2%。

     

    Abstract: Trypsin ⅠmRNA in two species of tilapia(Oreochromis aureus and Oreochromis niloticus) were cloned and sequenced by means of 3′/ 5′2 RACE.The results indicated that the size of trypsin ⅠmRNA of tilapia (O.niloticus) and tilapia (O.aureus) are 858bp and 863bp respectively,with an open reading frame of 738bp,which encoding trypsin containing 245 anmino acid residues with characteristic signal peptides and activation peptides.The tilapia trypsin Ⅰ have highly conserved amino acid residues essential for the catalytic activity and conformational maintenance,which are His,Asp,Ser of catalytic triad and twelve cysteines forming six disulfide bridges,together with the residues determining substrate specificity and generating the S1 substrate-binding pocket of a typical trypsin nature.The identity of trypsin ⅠmRNAof tilapia (O.aureus) compared with that of Antarctic fish (Patanotothenia magellanica),Antlantic Salmon ( Salmo salar),Flounder(Paralichthys olivaceus),Atlantic Cod(Gadusmorhua),Bovine,and Human was 58.3%—72.5 % and 63.3%—76.1%, respectively,And that in O.niloticus was59.1%—73.1% and 63.6%—75.6%.Trypsin I and its mRNA of O.niloticus showed identity of 96.2 % and 99.2% respectively to those of Oreochromis aureus.

     

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