期刊文献+

草鱼脂酰辅酶A合成酶4基因全长cDNA分子克隆及表达调控 预览

Full-Length cDNA Molecular Cloning and Expression Regulation of Acyl Coenzyme A Synthetase 4 Gene in Grass Carp
在线阅读 下载PDF
收藏 分享 导出
摘要 本试验采用反转录PCR(RT-PCR)技术克隆草鱼脂酰辅酶A合成酶4(ACSL4)基因全长cDNA,利用实时荧光定量PCR技术研究ACSL4基因在草鱼大脑、心脏、脾脏、肝胰脏、肌肉、肾脏、肠系膜脂肪、前肠、中肠、后肠10种组织中的表达情况,并对投喂不同淀粉源饲料以及饥饿再投喂0、3、6、12和24h后草鱼肝胰脏中ACSL4基因表达情况进行研究。结果显示:草鱼ACSL4基因cDNA全长2418bp,其开放阅读框2121bp,共编码706个氨基酸;草鱼ACSL4蛋白的氨基酸序列包含1个跨膜结构域、1个脂肪酸绑定基序和1个ATP/AMP绑定基序。草鱼ACSL4mRNA在大脑和脾脏中相对表达量较高,显著高于其他组织(P<0.05),在肌肉中相对表达量最低;投喂不同淀粉源饲料对草鱼肝胰脏中ACSL4mRNA相对表达量无显著影响(P>0.05);饥饿再投喂12h后,草鱼肝胰脏中ACSL4mRNA相对表达量达到最高值,显著高于其他时间段(P<0.05),24h后恢复至最初水平。本试验从草鱼10种混合组织中克隆了ACSL4基因全长cDNA,其所编码的氨基酸序列的主要功能性位点脂肪酸绑定基序和ATP/AMP绑定基序在不同物种间高度保守;草鱼ACSL4基因主要在大脑和脾脏中表达;饲料中淀粉源对草鱼肝胰脏中ACSL4基因的表达无显著影响;在饥饿再投喂12h后,草鱼肝胰脏中ACSL4mRNA相对表达量达到最高值,随后显著下降至最初水平。 The full-length cDNA of long chain acyl coenzyme A synthetase 4(ACSL4)gene was cloned from grass carp by reverse-transcription PCR(RT-PCR)technique.Tissue distribution of ACSL4 gene in the brain,heart,spleen,hepatopancreas,muscle,kidney,adipose tissues,anterior intestine,mid-intestine and posterior intestine of grass carp was analyzed by real-time quantitative PCR technique.Meanwhile,the ACSL4 gene expression in the hepatopancreas of grass carp fed with diets containing different starch sources was studied.In addition,the ACSL4 gene expression in the hepatopancreas of grass carp at 0,3,6,12 and 24 h after starvation and refeeding was also investigated.The results showed as follows:the full-length cDNA of ACSL4 gene was 2 418 bp with a 2 121 bp open reading frame(ORF)encoding 706 amino acids;the ACSL4 protein contained a trans-membrane domain,a ATP/AMP signature motif and a fatty acyl coenzyme A synthetase(FACS)signature motif.The relative expression level of ACSL4 mRNA in brain and spleen were higher,which were significantly higher than those in the other tissues(P<0.05),and the lowest in muscle.There were no significant differences of ACSL4 mRNA relative expression level in the hepatopancreas of grass carp after fed with diets containing different starch sources(P>0.05).However,the highest expression level of ACSL4 mRNA in the hepatopancreas of grass carp was detected after 12 h of starvation and refeeding,which was significantly higher than that at the other time(P<0.05),and then it returned to its beginning level after 24 h of starvation and refeeding.In summary,we have cloned the full-length cDNA of ACSL4 gene from grass carp,and the main functional sites ATP/AMP signature motif and FACS signature motif of amino acid sequences are both highly conserved in different species.Brain and spleen are the main ACSL4 gene expressing tissues in grass carp.Moreover,the ACSL4 gene expression in hepatopancreas of grass carp is not significantly affected by different dietary starch sources,and the highest relative expressi
作者 韩振 高琦 许建和 易乐飞 申欣 丁祝进 程汉良 HAN Zhen;GAO Qi;XU Jianhe;YI Lefei;SHEN Xin;DING Zhujin;CHENG Hanliang(College of Marine Life and Fisheries,Huaihai Institute of Technology,Lianyungang 222005,China)
机构地区 淮海工学院
出处 《动物营养学报》 CAS CSCD 北大核心 2019年第5期2442-2450,共9页 CHINESE JOURNAL OF ANIMAL NUTRITION
基金 国家自然科学基金项目(31272636) 江苏省自然科学基金项目(BK2012664) 江苏省优势学科建设工程项目(PAPD) 江苏省研究生科研与实践创新计划项目(SY201701X).
关键词 草鱼 长链脂酰辅酶A合成酶4 分子克隆 表达调控 grass carp long chain acyl coenzyme A synthetase 4 molecular cloning expression regulation
作者简介 韩振(1992-),男,江苏连云港人,硕士研究生,研究方向为水产动物营养与饲料。E-mail:513620638@qq.com;通信作者:程汉良,教授,硕士生导师,E-mail:CHL3139@163.com.
  • 相关文献

参考文献7

二级参考文献110

共引文献84

投稿分析

相关作者

内容加载中请稍等...

相关机构

内容加载中请稍等...

相关主题

内容加载中请稍等...

浏览历史

内容加载中请稍等...
;
使用帮助 返回顶部 意见反馈