| 香鱼甘油-3-磷酸脱氢酶基因的克隆与表达 |
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| 引用本文： | 郝晓娜,陈炯,陆新江,史雨红.香鱼甘油-3-磷酸脱氢酶基因的克隆与表达[J].生物技术通报,2011(2). |
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| 作者姓名： | 郝晓娜 陈炯 陆新江 史雨红 |
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| 作者单位： | 宁波大学应用海洋生物技术教育部重点实验室,宁波,315211 |
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| 基金项目： | 长江学者和创新团队发展计划项目(IRT0734) |
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| 摘 要： | GPDH(glycerol-3-phosphate dehydrogenase)是合成脂肪代谢中间产物甘油-3-磷酸的关键酶。通过设计简并引物从香鱼肝cDNA文库中克隆GPDH基因,该基因cDNA序列全长577个核苷酸,单一大的开放阅读框编码一个由351个氨基酸组成的分子量为37.9kD的蛋白。蛋白序列分析表明,香鱼GPDH(aGPDH)与亚洲胡瓜鱼的GPDH序列同源性最高。系统进化树分析表明,GPDH的物种进化关系与目前接受的物种分类关系基本一致。实时荧光定量PCR结果显示,aGPDH基因在香鱼肝、脾、肾、脑、心和肌肉组织均有表达。香鱼咸淡水适应以后,肝、脾、脑、心和肌肉的aGPDH的mRNA表达水平下调。成功构建重组表达质粒pET-32a-GPDH。SDS-PAGE试验表明,目的蛋白可以在大肠杆菌中大量表达;并制备了抗血清,能与目的蛋白起强的特异性反应,但不与细菌自身蛋白起反应。本研究有助于进一步理解鱼类盐度适应过程中的脂肪代谢调控机制。
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| 关 键 词： | GPDH 香鱼 基因克隆 盐度适应 实时荧光定量PCR 原核表达 |
cDNA Cloning and mRNA Expression of GPDH in Ayu(Plecoglossus altivelis) |
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| Hao Xiaona,Chen Jiong,Lu Xinjiang,Shi Yuhong.cDNA Cloning and mRNA Expression of GPDH in Ayu(Plecoglossus altivelis)[J].Biotechnology Bulletin,2011(2). |
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| Authors: | Hao Xiaona Chen Jiong Lu Xinjiang Shi Yuhong |
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| Institution: | Hao Xiaona Chen Jiong Lu Xinjiang Shi Yuhong(Key Laboratory of Applied Marine Biotechnology,Ministry of Education,Ningbo University,Ningbo 315211) |
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| Abstract: | Glycerol-3-phosphate dehydrogenase(GPDH),a key enzyme in liver catalyzes the reduction of dihydroxyacetone phosphate to glycerol-3-phosphate in lipid metabolism.In this study,a full-length cDNA clone of the GPDH gene,577 bp in size,encoding a protein of 351 amino acids with a molecular weight of 37.9 kD was isolated from the ayu(Plecoglossus altivelis)species.Sequence comparison showed that ayu GPDH(aGPDH)was most similar to rainbow smelt GPDH.The neighbour-joining tree showed that species of the same order... |
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| Keywords: | GPDH Ayu Gene cloning Salinity acclimation QRT-PCR Prokaryotic expression |
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