凡纳滨对虾ANT2基因的克隆及低温表达谱分析  

MOLECULAR CLONING OF LVANT2 GENE AND ITS EXPRESSION PATTERN BY COLD INDUCTION

作　　者：殷勤[1,2] 崔亮 彭金霞 韦嫔媛 谢达祥 陈秀荔 王志伟[2] 李奎[2] 陈晓汉 

机构地区：[1]广西壮族自治区水产研究所,广西水产遗传育种与健康养殖重点实验室,南宁530021 [2]中国农业科学院北京畜牧兽医研究所,北京100094

出　　处：《水生生物学报》2012年第1期24-28,共5页Acta Hydrobiologica Sinica

基　　金：广西科学研究与技术开发计划(桂科攻10100007-2);广西壮族自治区直属公益性科研院所基本科研业务费(2060302GXIF-2010-04);现代农业产业技术体系建设专项资金(编号:CARS-47)资助

摘　　要：为研究凡纳滨对虾适应低温的分子机理,实验对从低温处理凡纳滨对虾抑制性消减文库中筛选出的一个360 bp EST序列进行了研究。首先,同源比对显示该EST片段与其他物种的ANT2基因高度同源,命名为凡纳滨对虾ANT2基因(LVANT2);其次,通过构建凡纳滨对虾肝胰腺全长cDNA文库,PCR扩增获得LVANT2基因的全长cDNA1540 bp,其中包括1011 bp的完整开放阅读框,编码336个氨基酸残基。然后,对基因进行了不同组织和低温处理的表达谱分析:(1)组织表达谱的结果显示,该基因在凡纳滨对虾肌肉组织中表达量最高;(2)在不同低温处理下的表达结果显示,该基因在15℃处理下基因表达量发生显著变化,13℃开始呈下调表达,11℃时表达量又升高;13℃低温处理不同时间发现该基因在12h内表达量发生显著变化,48h后表达量最高。LVANT2基因的低温诱导表达模式说明其可能在凡纳滨对虾低温适应中发挥作用。To study the molecular mechanism of cold tolerance of Litopenaeusvannamei,a 360 bp EST was isolated from suppression subtractive hybridization library of hepatopancreas which treated by low temperature.The 360 bp EST shared highly homology with ANT2 gene of other species,so we named it Litopenaeus vannamei ANT2 gene（LVANT2）.Then we cloned whole cDNA sequence of PVANT2 from a hepatopancreas cDNA library.The LVANT2 gene was 1540 bp with a 1011 bp open reading frame encoding a peptide of 309aa.Tissue expression pattern by real-time PCR revealed that LVANT2 mRNA existed in heart,hepatopancreas,muscle,stomach,intestinal and eyestalk but mainly existed in muscle tissue.Cold induction by different temperatures revealed that LVANT2 mRNA expression started to increase after 15℃ treatment for 36 hours,and then decreased after 13℃ treatment for 36 hours,but increased again after 11℃ treatment.As for 13℃ treatment for different times,the expression pattern changed during the first 12 hours,but the expression with low temperature treatment for 24 hours and 36 hours was almost the same with the room temperature.The low temperature induction expression pattern demonstrated that LVANT2 may function in cold tolerance of Litopenaeus vannamei.

关 键 词：LVANT2 表达量 抑制性消减文库 cDNA全长文库 

分 类 号：Q781[生物学—分子生物学]