金纹细蛾β-actin基因克隆及表达分析  

Cloning and Expression Analysis of β-actin Gene from Phyllonorycter ringoniellan(Lepidoptera:Gracilariidae)

作　　者：郭长宁[1] 李鑫[1] 于建光[1] 侯茜[1] 

机构地区：[1]西北农林科技大学植物保护学院,应用昆虫学重点实验室,陕西杨凌712100

出　　处：《西北农业学报》2015年第3期157-162,共6页Acta Agriculturae Boreali-occidentalis Sinica

基　　金：陕西省农业攻关(2008K01-04);西北农林科技大学科技创新与成果转化(XNY2013-67)

摘　　要：为了明确β-actin基因在金纹细蛾定量实验中作为内参基因的可靠性。根据近缘物种同源基因设计简并引物,通过RT-PCR技术克隆得到金纹细蛾β-actin基因片段,并进行序列分析;利用QRT-PCR方法检测该基因在金纹细蛾不同发育时期及成虫5种组织间的表达情况。结果表明:金纹细蛾β-actin基因片段为594bp(GenBank登录号:KF880733),编码174个氨基酸残基,具有actin蛋白家族典型特征,富含4种类型特定功能位点,其氨基酸序列与其他鳞翅目昆虫的相似性均大于96%。金纹细蛾β-actin基因在其生长发育阶段稳定表达,无显著差异,可以作为研究金纹细蛾不同发育时期的内参;在成虫头、胸、足和翅4种组织间表达量无显著差异。The purpose of this research is to make clear the reliability ofβ-actin as reference gene in quantitative experiments from Phyllonorycter ringoniellan.Degenerate primers were designed based on homologous gene from other species,theβ-actin gene from P.ringoniella was cloned.Moreover,the QRT-PCR was performed to detect the gene expression levels in developmental stages and five different adult tissues（head,thorax,abdomen,leg and wing）.Sequence analysis indicated thatβ-actin gene fragment（KF880733）was 594 bp,encoding 174 amino acid residues.The deduced protein contained 4functional sites and was above 96%identical withβ-actin genes from other Lepidoptera insects.Quantitative assays revealed that there was no significant difference between either different developmental stages or four adult tissues（head,thorax,leg and wng）.The result indicated thatβ-actin gene is a reliable internal control in the developmental stages of P.ringoniella.

关 键 词：金纹细蛾 Β-ACTIN基因 序列分析 基因表达 

分 类 号：S433.4[农业科学—农业昆虫与害虫防治]