凡纳滨对虾烯醇酶基因Enolase的克隆及表达分析  

Clone and expression analysis of Enolase in Litopenaeus vannamei

作　　者：李文兰[1] 高永华[1,2] 盛小伟[1,2] 陈晓汉[2] 熊建华[2] 

机构地区：[1]桂林理工大学化学与生物工程学院,广西桂林541000 [2]广西水产研究所南美白对虾遗传育种中心,广西南宁530021

出　　处：《广东农业科学》2011年第5期168-171,174,共5页Guangdong Agricultural Sciences

基　　金：国家科技支撑计划项目(2008BADB9B03);广西直属公益性科研院所基本科研业务费(2060302GXIF-2008-02);国家虾产业技术体系岗位科学教经费(nycytx-46)

摘　　要：以生长性状分离的凡纳滨对虾家系的肌肉组织,构建凡纳滨对虾生长性状差减cDNA文库,斑点杂交筛选获得Enolase基因,克隆获得全长编码区序列。序列分析表明,序列包含1 305 bp的开放阅读框,推测编码的蛋白质含434个氨基酸,预测的分子量为47.4 ku,等电点为5.67。通过荧光定量PCR法研究了Enolase基因在凡纳滨对虾的不同组织和不同生长期的肌肉组织中的表达情况,结果显示,Enolase基因在凡纳滨对虾的心脏、肝胰腺、肌肉、胃、肠、眼柄、鳃和血等组织或器官中都有表达;在肌肉组织中表达量最高,在肝胰腺中表达量最低;在日龄为125 d的对虾肌肉组织中表达量最高,在日龄为80 d的对虾肌肉组织中表达量最低。A subtracted cDNA libraries of Litopenaeus vannamei on growth traits was constructed,and enolase gene of Litopenaeus vannamei was gained by dot blot and cloning.The sequence contained a 1 305 bp open reading frame（ORF） encoding 434 amino acids（aa）.The predicted molecular weight was 47.4 ku and the theoretical isoelectric point was 5.67.Quantitative real-time PCR was used to assess the mRNA expression of enolase in different tissue and different growth period.Its expression was detected in heart,hepatopancreas,muscles,stomach,bowel,eye hilt,gill and blood.The highest expression level was observed in muscle,while the lowest in hepatopancreas;the expression of enolase in different growth showed that the highest expression level in muscles was 125 days age,while the lowest was 80 days age.

关 键 词：凡纳滨对虾 烯醇酶 抑制性差减杂交 荧光定量PCR 

分 类 号：S966.12[农业科学—水产养殖]