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首页> 外文期刊>Aquaculture >Comparison of intestinal gene expression in atlantic cod (Gadus morhua) fed standard fish meal or soybean meal by means of suppression subtractive hybridization and real-time PCR.
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Comparison of intestinal gene expression in atlantic cod (Gadus morhua) fed standard fish meal or soybean meal by means of suppression subtractive hybridization and real-time PCR.

机译:通过抑制消减杂交和实时荧光定量PCR(PCR)比较大西洋鳕鱼(Gadus morhua)饲喂标准鱼粉或豆粕中肠道基因的表达。

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摘要

Gene expression was studied in Atlantic cod fed two different diets, fish meal (FM) and dehulled and extracted soybean meal (SBM). RNA was isolated from the distal part of the mid-intestine of Atlantic cod and suppression subtractive hybridization (SSH) was employed to screen for genes that showed changes in expression in response to the two dietary treatments. We made a cDNA subtracted library, isolated and sequenced 192 clones. Identification of 157 clones was predicted by BLAST. Most of the clones were previously unidentified in cod. Expression of 12 selected clones was further studied by quantitative PCR. Expression of four clones showing similarity to aminopeptidase N, transcobalamin I precursor, cytochrome P450 3A40, and ras-related nuclear protein was significantly up regulated in intestine of cod fed SBM compared to cod fed FM. A trend towards up regulation of a clone with similarity to fatty acid binding protein in SBM-fed cod was also observed. No significant differences in expression were observed for: transmembrane 4 superfamily protein member, polypeptide N-acetylgalactosaminyltransferase, glutathione peroxidase, peroxiredoxin 4, SEC61, F-BOX, and 14-3-3..
机译:在大西洋鳕鱼中,研究了两种鱼粮(鱼粉(FM))和脱壳和提取的豆粕(SBM)的基因表达。从大西洋鳕鱼肠中段的远端分离RNA,并采用抑制消减杂交(SSH)来筛选基因,这些基因在两种饮食疗法中均表现出表达变化。我们制作了一个cDNA减去文库,分离并测序了192个克隆。通过BLAST预测了157个克隆的鉴定。大多数克隆以前未在鳕鱼中鉴定。通过定量PCR进一步研究了12种选择的克隆的表达。与鳕鱼喂养的FM相比,鳕鱼喂养的SBM的肠中四个与氨肽酶N,反钴胺素I前体,细胞色素P450 3A40和ras相关的核蛋白相似的克隆的表达明显上调。还观察到了由SBM喂养的鳕鱼中与脂肪酸结合蛋白相似的克隆的上调趋势。没有观察到表达的显着差异:跨膜4超家族蛋白成员,多肽N-乙酰半乳糖胺基转移酶,谷胱甘肽过氧化物酶,过氧化物酶4,SEC61,F-BOX和14-3-3。

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