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三疣梭子蟹血细胞全长cDNA文库构建及EST初步分析
引用本文:申望,叶茂,石戈,王日昕.三疣梭子蟹血细胞全长cDNA文库构建及EST初步分析[J].海洋渔业,2010,32(1):24-29.
作者姓名:申望  叶茂  石戈  王日昕
作者单位:浙江海洋学院海洋科学学院,海洋生物资源及分子工程实验室,舟山,316004
基金项目:浙江省重大科技专项重大项目(2007C02001);;浙江省教育厅项目(20061141)
摘    要:三疣梭子蟹是我国沿海重要养殖品种之一,近年来养殖病害呈逐年上升趋势,制约了三疣梭子蟹养殖产业的健康可持续发展。为大规模克隆三疣梭子蟹免疫相关基因,研究其免疫基因的功能和免疫机制,奠定三疣梭子蟹病害防治理论基础,采用SMART技术构建了三疣梭子蟹血细胞全长cDNA文库。经测定,文库滴度为1.14×107pfu/mL,文库总容量为5.7×107pfu,重组率达到98%,插入片段平均长度为817.5 bp。文库随机测序获得的70个全长cDNA,从中发现18个已知基因和5个未知基因。已知基因主要为多肽/蛋白质合成相关基因,共9个;免疫相关基因4个,分别是抗菌肽hyastatin、C-reactive prote in-1、profilin和m asquerade-like prote in,且4个免疫相关基因共有48个克隆,占测序总克隆的68.6%,表明血细胞中免疫相关基因表达非常活跃。研究结果证实,构建血细胞全长cDNA文库是大规模克隆三疣梭子蟹免疫相关基因的可靠途径,并为深入研究免疫相关基因的功能和免疫机制奠定基础。

关 键 词:三疣梭子蟹  血细胞  全长cDNA文库  免疫相关基因  表达序列标签

Construction of a full-length cDNA library of Portunus trituberculatus hemocytes and analysis of partial expressed sequence tags
SHEN Wang,YE Mao,SHI Ge,WANG Ri-xin.Construction of a full-length cDNA library of Portunus trituberculatus hemocytes and analysis of partial expressed sequence tags[J].Marine Fisheries,2010,32(1):24-29.
Authors:SHEN Wang  YE Mao  SHI Ge  WANG Ri-xin
Institution:Laboratory of Marine Biology Resources and Molecular Engineering;Ocean Science College;Zhejiang Ocean University;Zhoushan 316004;China
Abstract:Portunus trituberculatus is one of the most important aquaculture crustacean species in China.But the sustainable development of P.trituberculatus aquaculture had been restricted by ascendant aquatic disease.And the immune system of P.trituberculatus has always been an hot issue to researchers.The immunity of P.trituberculatus is related to various immune molecules in plasma.To obtain the full-length cDNA sequences of potential immune-related genes and explore their function,a high-quality cDNA library of P...
Keywords:Portunus trituberculatus  hemocyte  full-length cDNA library  immune-related genes  ESTs  
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