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半滑舌鳎AKT-ineractingprotein基因的克隆及免疫应答表达分析
  • ISSN号:1000-3207
  • 期刊名称:《水生生物学报》
  • 时间:0
  • 分类:Q522[生物学—生物化学]
  • 作者机构:[1]大连海洋大学水产与生命学院,大连116023, [2]中国水产科学研究院黄海水产研究所农业部海洋渔业可持续发展重点实验室,青岛266071, [3]青岛海洋科学与技术国家实验室海洋渔业科学与食物产出过程功能实验室,青岛266200, [4]上海海洋大学水产与生命学院,上海201306
  • 相关基金:国家“863”高技术研究发展计划(2012AA10A401-4);国家自然科学基金(31172439)资助
中文摘要:

研究克隆了半滑舌鳎Cynoglossussemilaevis膜蛋白AKT—interactingprotein(AKTIP)基因,研究了其在健康组织中的表达模式、鳗弧菌(Vibrioanguillarum)感染后免疫组织和不同病原物刺激外周血淋巴细胞中的表达特征。通过常规克隆和RACE技术,获得的半滑舌鳎AKTIP基因全长cDNA序列为1224bp,其中包括5’-UTR为116bp,3'-UTR为117bp和完整的ORF序列891bp,编码296个氨基酸,预测蛋白质的等电点(PI)为9.12,分子量是33.74kD;同源比对发现半滑舌鳎AKTIP的氨基酸序列在不同物种之间具有较高的保守性;荧光实时定量PCR(qRT-PCR)检测到半滑舌鳎各个组织中均有4KTIP基因的表达,在卵巢中表达量最高;鳗弧菌感染半滑舌鳎后,AKTIP基因在肝、鳃、血液、肠、头肾和脾中均上调表达;病原模拟物PGN、LPS、polyI:C和WGP刺激半滑舌鳎外周血淋巴细胞均诱导AKTIP基因下调表达。研究表明半滑舌鳎AKTIP基因参与了机体免疫反应,为给半滑舌鳎免疫防御技术的研究提供理论依据。

英文摘要:

AKT-interacting protien (AKTIP) is a kind of membrane protein, involvimg in the regulation of PI3K/PDK1/Akt pathway. AKTIP gene is still unknown in fish. In the present study, AKTIP gene of Cynoglossus semilaevis was first cloned and full length of cDNA was 1224 bp in size, including 5'-untranslated region (UTR) of 116 bp, 3'-UTR of 117 bp and a complete open reading frame (ORF) of 891 bp, encoding 296 amino acids. Theoretical isoelectric point (PI) of predicted protein was 9.12 and molecular weight was 33.74 kD. Homologous comparison showed that the amino acids sequence of AKTIP in C. semilaevis had a high identity with those of other species. The AKTIP gene was expressed in all tested tissues in the health C. semilaevis and the highest expression was in the ovary. In order to investigate the expression patterns of the AKTIP gene in immune response, the specific expression of AKTIP was performed after Vibrio anguillarum infection and in peripheral blood lymphocytes with different pathogens stimulated. The results showed that the expression ofAKTIP gene was up-regulated in the liver, gill, blood, intestinal, head kidney and spleen after V. anguillarum infection. While the AKTIP gene expression in peripheral blood lymphocytes was down-regulated after PGN, LPS, WGP and poly I: C stimulation. The research revealed that AKTIP gene involved in C. semilaevis immune response. These results provide the evidence to explore AKTIP defense mechanism and to control C. semilaevis disease in the further study.

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期刊信息
  • 《水生生物学报》
  • 北大核心期刊(2011版)
  • 主管单位:中国科学院
  • 主办单位:中科院水生所
  • 主编:桂建芳
  • 地址:武昌东湖南路7号中科院水生所
  • 邮编:430072
  • 邮箱:Acta@ihb.ac.cn
  • 电话:027-68780701
  • 国际标准刊号:ISSN:1000-3207
  • 国内统一刊号:ISSN:42-1230/Q
  • 邮发代号:82-329
  • 获奖情况:
  • 湖北省十佳重点期刊,中国农学会中国水产学会优秀科技期刊一等奖,中国科学院优秀期刊三等奖,中国期刊方阵“双效”期刊
  • 国内外数据库收录:
  • 美国化学文摘(网络版),英国农业与生物科学研究中心文摘,美国生物科学数据库,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),中国北大核心期刊(2000版)
  • 被引量:21674