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A key gene of the RNA interference pathway in the black tiger shrimp, Penaeus monodon: Identification and functional characterisation of Dicer-1

文献类型：期刊论文

作者Su, Jianguo1,2,3,4; Oanh, Dang T. H.1,2; Lyons, Russell E.1,2; Leeton, Lisa1,2; van Hulten, Marielle C. W.1,2; Tan, Siok-Hwee1,2; Song, Linsheng4; Rajendran, K. V.1,2; Walker, Peter J.1,2
刊名FISH & SHELLFISH IMMUNOLOGY
出版日期2008-02-01
卷号24期号:2页码:223-233
关键词Penaeus Monodon Dicer Rna Interference Gene Cloning Characterisation
ISSN号1050-4648
DOI10.1016/j.fsi.2007.11.006
文献子类Article
英文摘要RNA interference (RNAi) is an evolutionarily conserved mechanism by which double-stranded RNA (dsRNA) initiates post-transcriptional silencing of homologous genes. Here we report the amplification and characterisation of a full length cDNA from black tiger shrimp (Penaeus monodon) that encodes the bidentate RNAase III Dicer, a key component of the RNAi pathway. The full length of the shrimp Dicer (Pm Dcr1) cDNA is 7629 bp in length, including a 51 untranslated region (UTR) of 130 bp, a 3' UTR of 77 bp, and an open reading frame of 7422 bp encoding a polypeptide of 2473 amino acids with an estimated molecular mass of 277.895 kDa and a predicted isoelectric point of 4.86. Analysis of the deduced amino acid sequence indicated that the mature peptide contains all the seven recognised functional domains and is most similar to the mosquito (Aedes aegypti) Dicer-1 sequence with a similarity of 34.6%. Quantitative RT-PCR analysis showed that Pm Dcr1 mRNA is most highly expressed in haemolymph and lymphoid organ tissues (P 0.05). However, there was no correlation between Pm Dcr1 mRNA levels in lymphoid organ and the viral genetic loads in shrimp naturally infected with gill-associated virus (GAV) and Mourilyan virus (P > 0.05). Treatment with synthetic dsRNA corresponding to Pm Dcr1 sequence resulted in knock-down of Pm Dcr1 mRNA expression in both uninfected shrimp and shrimp infected experimentally with GAV. Knock-down of Pm Dcr1 expression resulted in more rapid mortalities and higher viral loads. These data demonstrated that Dicer is involved in antiviral defence in shrimp. (c) 2007 Elsevier Ltd. All rights reserved.; RNA interference (RNAi) is an evolutionarily conserved mechanism by which double-stranded RNA (dsRNA) initiates post-transcriptional silencing of homologous genes. Here we report the amplification and characterisation of a full length cDNA from black tiger shrimp (Penaeus monodon) that encodes the bidentate RNAase III Dicer, a key component of the RNAi pathway. The full length of the shrimp Dicer (Pm Dcr1) cDNA is 7629 bp in length, including a 51 untranslated region (UTR) of 130 bp, a 3' UTR of 77 bp, and an open reading frame of 7422 bp encoding a polypeptide of 2473 amino acids with an estimated molecular mass of 277.895 kDa and a predicted isoelectric point of 4.86. Analysis of the deduced amino acid sequence indicated that the mature peptide contains all the seven recognised functional domains and is most similar to the mosquito (Aedes aegypti) Dicer-1 sequence with a similarity of 34.6%. Quantitative RT-PCR analysis showed that Pm Dcr1 mRNA is most highly expressed in haemolymph and lymphoid organ tissues (P 0.05). However, there was no correlation between Pm Dcr1 mRNA levels in lymphoid organ and the viral genetic loads in shrimp naturally infected with gill-associated virus (GAV) and Mourilyan virus (P > 0.05). Treatment with synthetic dsRNA corresponding to Pm Dcr1 sequence resulted in knock-down of Pm Dcr1 mRNA expression in both uninfected shrimp and shrimp infected experimentally with GAV. Knock-down of Pm Dcr1 expression resulted in more rapid mortalities and higher viral loads. These data demonstrated that Dicer is involved in antiviral defence in shrimp. (c) 2007 Elsevier Ltd. All rights reserved.
学科主题Fisheries ; Immunology ; Marine & Freshwater Biology ; Veterinary Sciences
URL标识查看原文
语种英语
WOS记录号WOS:000253402200010
公开日期2010-12-24
源URL[http://ir.qdio.ac.cn/handle/337002/5643]  
专题海洋研究所_实验海洋生物学重点实验室
作者单位1.CSIRO Livestock Ind, Queensland Biosci Precinct, Geelong, Vic 3220, Australia
2.CSIRO Livestock Ind, Australian Anim Hlth Lab, Geelong, Vic 3220, Australia
3.NW A&F Univ, Coll Anim Sci, Dept Aquaculture, Yangling 712100, Peoples R China
4.Chinese Acad Sci, EMBL, Inst Oceanol, Qingdao 266071, Peoples R China
推荐引用方式
GB/T 7714		 Su, Jianguo,Oanh, Dang T. H.,Lyons, Russell E.,et al. A key gene of the RNA interference pathway in the black tiger shrimp, Penaeus monodon: Identification and functional characterisation of Dicer-1[J]. FISH & SHELLFISH IMMUNOLOGY,2008,24(2):223-233.
APA Su, Jianguo.,Oanh, Dang T. H..,Lyons, Russell E..,Leeton, Lisa.,van Hulten, Marielle C. W..,...&Walker, Peter J..(2008).A key gene of the RNA interference pathway in the black tiger shrimp, Penaeus monodon: Identification and functional characterisation of Dicer-1.FISH & SHELLFISH IMMUNOLOGY,24(2),223-233.
MLA Su, Jianguo,et al."A key gene of the RNA interference pathway in the black tiger shrimp, Penaeus monodon: Identification and functional characterisation of Dicer-1".FISH & SHELLFISH IMMUNOLOGY 24.2(2008):223-233.

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来源：海洋研究所

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