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草鱼 Ctenopharyngodon idella TLR7 的鉴定及对双链 RNA 和病毒感染的表达谱分析。

Identification and expression profiles of grass carp Ctenopharyngodon idella tlr7 in responses to double-stranded RNA and virus infection.

机构信息

College of Animal Science and Technology College of Veterinary Medicine Northwest A&F University, Shaanxi Key Laboratory of Molecular Biology for Agriculture, Yangling 712100, China.

出版信息

J Fish Biol. 2012 Jun;80(7):2605-22. doi: 10.1111/j.1095-8649.2012.03316.x.

Abstract

The toll-like receptor 7 (tlr7) gene was identified and characterized from grass carp Ctenopharyngodon idella (designated as citlr7), and the messenger (m)RNA expression profiles were examined in vivo and in vitro. The citlr7 genomic sequence consists of 4276 nucleotides (nts), including two exons and one intron. The full length of citlr7 complementary (c)DNA sequence is 3354 nts with the longest open reading frame (ORF) of 3156 nts encoding a peptide of 1051 amino acids. citlr7 mRNA expression was high in spleen, skin and heart, and low in hepatopancreas, muscle, head kidney and trunk kidney in healthy fish. The expression of citlr7 was rapidly and significantly up-regulated at 6 h after grass carp reovirus (GCRV) injection (72·91 fold, P < 0·05), and recovered to the original level at 24 h post-injection in the spleen. The citlr7 transcript was rapidly and significantly down-regulated at the 6 h time point (0·32 fold, P < 0·05) and retrieved the normal level at 72 h post-injection in the hepatopancreas. The citlr7 transcripts were rapidly and significantly inhibited at 2 h post-GCRV infection in the C. idella kidney (CIK) cell line (0·62 fold, P < 0·05), and were rapidly and significantly elevated by the stimulation of the synthetic double-stranded RNA polyriboinosinic-polyribocytidylic acid sodium salt [poly(I:C)] in CIK in a dose and time-dependent manner (P < 0·05). The results imply that citlr7 is involved in the responses to double-stranded RNA and virus infection.

摘要

从草鱼(Ctenopharyngodon idella)中鉴定和表征了 Toll 样受体 7(tlr7)基因(命名为 citlr7),并在体内和体外检查了信使(m)RNA 的表达谱。citlr7 基因组序列由 4276 个核苷酸(nts)组成,包括两个外显子和一个内含子。citlr7 全长互补(c)DNA 序列为 3354 nts,最长开放阅读框(ORF)为 3156 nts,编码 1051 个氨基酸的肽。在健康鱼类中,citlr7 mRNA 在脾脏、皮肤和心脏中的表达量较高,在肝胰腺、肌肉、头肾和中肾中的表达量较低。草鱼呼肠孤病毒(GCRV)注射后 6 h,citlr7 的表达迅速显著上调(72·91 倍,P < 0·05),脾脏注射后 24 h 恢复到原始水平。注射后 6 h,肝胰腺 citlr7 转录物迅速显著下调(0·32 倍,P < 0·05),72 h 恢复正常水平。在草鱼肾脏(CIK)细胞系中,GCRV 感染后 2 h,citlr7 转录物迅速显著抑制(0·62 倍,P < 0·05),用聚肌苷酸-聚胞苷酸钠盐[poly(I:C)]刺激后迅速显著升高,呈剂量和时间依赖性(P < 0·05)。结果表明 citlr7 参与了对双链 RNA 和病毒感染的反应。

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