谢氏宽漠王β-actin基因cDNA克隆、序列分析及表达量检测  

Cloning,characterization and expression detection of the β-actin cDNA from Mantichorula semenowi (Coleoptera:Tenebrionidae)

作　　者：唐婷[1] 柳峰松[1] 任国栋[1] 

机构地区：[1]河北大学生命科学学院,河北保定071002

出　　处：《昆虫学报》2008年第11期1210-1215,共6页Acta Entomologica Sinica

基　　金：国家自然科学基金重点项目(30130040);河北省无脊椎动物系统学与应用实验室资助项目(HBI08001)

摘　　要：β-actin是actin家族的一员,在维持细胞结构、运动和分裂等细胞生理活动方面发挥着重要作用,是基因定量实验中最常用的内参之一。本实验采用同源克隆和RACE技术扩增得到谢氏宽漠王Mantichorula semenowi β-actin基因。序列分析结果表明,该基因cDNA全长1372bp,开放阅读框长1131bp,编码376个氨基酸,5′和3′末端非翻译区域(UTR)分别为66bp和175bp;该序列与其他动物β-actin基因核苷酸序列具有96%～99%高度同源性。β-actin表达量检测结果显示热激后不同恢复时间其表达量无明显变化,且与未经热激处理的对照相比无显著差异。表明β-actin是研究受外界环境胁迫作用下昆虫体内不同基因表达水平的可靠内参基因。β-actin is a member of the actin family, which plays important roles in maintaining cytoskeletal structure, cytoplasmic movement and cell division, β-actin is one of the most commonly used internal controls in gene quantitative analysis. In this study, a full-length cDNA of β-actin from Mantichorula semenowi was cloned via homology cloning and RACE. Sequence analysis and phylogenetie study showed that this cDNA contained 1 372 nucleotides, with a 1 131 bp open reading frame （ORF） encoding 376 amino acids and flanked with a 66 bp 5＇UTR and a 175 bp 3＇UTR. β-actin shared a high sequence identity of 96% - 99% with that of other insects. Profiles of β-actin expression in different recovery time after heat shock were expressed similarly and not significantly different from that of the unstimulated control, suggesting an ubiguitous expression pattern.

关 键 词：谢氏宽漠王 Β-ACTIN基因 同源克隆 cDNA快速末端扩增 分子内标 

分 类 号：Q966[生物学—昆虫学]