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低盐胁迫下凡纳滨对虾鳃丝抑制性消减杂交cDNA文库构建及差异ESTs分析
  • ISSN号:1003-6482
  • 期刊名称:《海洋湖沼通报》
  • 时间:0
  • 分类:S917.4[农业科学—水产科学]
  • 作者机构:[1]广东海洋大学水产学院水产动物营养与饲料实验室,广东湛江524025
  • 相关基金:本研究由国家自然科学基金(30871928)、广东省自然科学基金(9152408801000006)、广东省“千百十”工程国家级人才培养项目及广东省“珠江学者”GDUPS(2011)项目资助
中文摘要:

本研究以几纳滨对虾(Lito penaeusvunnmei)为对象.分别以低盐度胁迫下(4ppt)凡纳滨对虾鳃丝cDNA为检测予,正常海水中凡纳滨对虾鳃丝cDNA为驱动子,采用抑制性消减杂交(suppressionsubtractive hybridization,SSH)技术构建了急性低盐胁迫诱导下凡纳滨对虾鳃丝消减cDNA文库。文库的重组率均高于95%,插入片段集中在250~750bp之间,随机测序后在线拼接得到15组片段重叠群和37个单一序列共j2个非重复序列。同源检索发现30个非重复序列与GenBank中的已知基因序列有较高的相似性。按差异表达基因编码的蛋白具体功能可分为6小类.依次为:(1)离子通道及转运蛋白;(2)蛋白合成翻译及转录因子;(3)应激抗氧化因子;(4)能量代谢;(5)信号受体和转导;(6)细胞纤维及骨架蛋白,所占比例分别为23.3%、20.0%、20.0%、16.7%、10.0%和10.0%。经功能聚类分析发现文库中舍有大量离子通道蛋白及功能转运酶、胞内信号传导分子、细胞骨架蛋白等渗透调节和应激适应性相关基因,表明急性低盐胁迫诱导几纳滨鳃丝抑制性消减杂交cDNA文库构建成功。

英文摘要:

A suppression subtractive hybridization (SSH) cDNA library of postgills in Litopenaeus van- ,amei induced by acute low salinity stress was constructed in this study. The postgills of L. vannamei in hypo-osmotic stress (4 ppt) were used as the monitoring and those in normal seawater (31 ppt) as the driver element, cDNA clones were selected from library and then sequenced. These sequences were analyzed using BLASTX software and classed for the results. The recombinant efficiency of cDNA li- brary was higher than 95o/00 and the inserted fragment was concentrated between 250~750 bp. Total 81 expressed sequence tags (ESTs) were obtained by randomly sequencing, and 52 unigenes including 15 contigs and 37 singletons after CP3 online matching were gained by clustering and assembling. The re sults of sequence homology search showed that there were 80 known with significant homology to Gen- bank database sequences that were identified, while 22 unigenes did not match any sequences in the datebase and presumably represented unidentified cDNAs. The clusters were concluded into 6 catego- ries according to their translated protein functions. They were ion channels and transport proteins (1) protein synthesis, translation and transcription factor (2) stress resistance and antioxidant factors (3) energy metabolism (4) signal receptor and transduction (5) cell fibrin and cytoskeletal proteins (6) which are account for 23.3%, 20.0%, 20.0%, 16.7%, 10.0%, and 10.0% respectively. A lot of os- moregulation ralated genes were discovered and identified and the SSH cDNA library of postgills in Litoperzaeus vannamei induced by acute low salinity stress were constructed successfully.

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期刊信息
  • 《海洋湖沼通报》
  • 北大核心期刊(2011版)
  • 主管单位:山东省科学技术协会
  • 主办单位:山东海洋湖沼学会
  • 主编:赵栋梁
  • 地址:青岛市鱼山路五号
  • 邮编:266003
  • 邮箱:
  • 电话:0532-82032650
  • 国际标准刊号:ISSN:1003-6482
  • 国内统一刊号:ISSN:37-1141/P
  • 邮发代号:24-83
  • 获奖情况:
  • 国内外数据库收录:
  • 美国剑桥科学文摘,英国动物学记录,日本日本科学技术振兴机构数据库,中国中国科技核心期刊,中国北大核心期刊(2004版),中国北大核心期刊(2008版),中国北大核心期刊(2011版),中国北大核心期刊(2014版)
  • 被引量:7653