中国水产科学2011，Vol.18Issue(1)：38-47,10.DOI:10.3724/SP.J.1118.2011.00038
草鱼天然抗性相关巨噬蛋白基因全长cDNA的克隆与表达分析
Molecular cloning, characterization and expression of natural resistance associated macrophage protein(Nramp)gene cDNA from grass carp(Ctenopharyngodon idella)
摘要
Abstract
Natural resistance associated macrophage protein (Nramp) family is a novel family of functional related proteins with innate resistance to intracellular parasites in mammals, which has the capacity of enhancing macrophages that are meant to kill pathogens. However, few reports are available about the structure and function of Nramp in lower vertebrates such as teleosts. In this study, cDNA of Nramp gene was amplified from liver of grass carp (Ctenopharyngodon idella) by reverse transcription polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE). The full-length cDNA of Nramp gene was 3 158 bp, including a 155 bp 5'terminal UTR, a 1 368 bp 3' terminal UTR and a 1 635 bp open reading frame encoding a protein with 554 amino acid residues, which contained the signature features of the Nramp protein family: 12 transmembrane (TM)domains, a consensus transport motif (CTM) with 20 amino acid residues between the transmembrane domains 8 and 9. The deduced amino acid sequence of grass carp Nramp exhibited about 62.5％-90.2％ homology with 16 other vertebrate Nramp sequences. Compared with the other fish's Nramps, the unique feature of grass carp Nramp was the presence of two iron-responsive regulatory-protein-binding sites (IRE) in 3'UTR and 5'UTR. Phylogenetic analysis revealed that the grass carp Nramp was clustered with other fish Nramps and was closer to Nramp2 of mammals. RT-PCR results of grass carp Nramp transcripts in different tissues indicated that the grass carp Nramp transcripts were highly abundant in head kidney and spleen and low in muscle and skin. The grass carp kidney cells (CIK) challenged with the grass carp reovirus (GCRV) could evidently elevate Nramp mRNA levels in cell cultures.关键词
草鱼/天然抗性巨噬蛋白基因(Nramp)/cDNA/表达/RT-PCR/RACE分类
生物科学引用本文复制引用
范玉顶,徐进,罗晓松,周勇,肖艺,曾令兵..草鱼天然抗性相关巨噬蛋白基因全长cDNA的克隆与表达分析[J].中国水产科学,2011,18(1):38-47,10.基金项目
农业部现代农业产业技术体系建设专项资金项目(nycytx-49-16) （nycytx-49-16）
中央级公益性科研院所基本科研业务费专项资金项目(2008JBYWF-22) （2008JBYWF-22）
长江水产研究所所长基金项目(SZ2008-05). （SZ2008-05）
