橘小实蝇α1微管蛋白基因的克隆和表达分析  

Cloning and expression pattern analysis of α1-tubulin in Bactrocera dorsalis(Hendel)

作　　者：申建梅[1] 胡黎明[1] 宾淑英[1] 廖泓之[1] 林进添[1] 

机构地区：[1]仲恺农业工程学院/外来有害生物预警与控制研究所,广东广州510225

出　　处：《南京农业大学学报》2011年第4期48-54,共7页Journal of Nanjing Agricultural University

基　　金：国家自然科学基金项目(30971923);仲恺农业工程学院科研项目(G3100003)

摘　　要：利用RT-PCR和RACE方法克隆获得橘小实蝇α1微管蛋白基因的cDNA全长序列,命名为α1-Bdortub。测序结果表明,α1-Bdortub开放阅读框全长1 353 bp,编码450个氨基酸残基。氨基酸序列比对表明,α1-Bdortub与黑腹果蝇α1-tubulin的氨基酸序列相似性最高,为99.8%。不同部位和不同时期的实时荧光定量PCR分析表明,α1-Bdortub在不同部位都有表达,雌虫翅中的表达是其生殖节中的12倍。从幼虫期到蛹期α1-Bdortub mRNA表达量逐渐提高;第1、2、3天幼虫中的含量分别为第10天蛹的0.71、1.16、4.01倍;在第1、4、7天蛹中的含量分别为第10天蛹的4.22、13.2、18.5倍。结果提示,α1-Bdortub可能与橘小实蝇的发育有关。α1-tubulin was cloned from Bactrocera dorsalis and named as α1-Bdortub.The sequencing results showed that the full-length of open reading frame in α1-Bdortub is 1 353 bp,encoding 450 amino acid residues.The putative amino acid shared the highest similarity（99.8%）with α1-tubulin from Drosophila melanogaster.Tissue expression revealed that α1-Bdortub expressed in all tissues and the relative expression level in female wings is about 12 folds of that in female genital segment.The temporal pattern further indicated that α1-Bdortub mRNA was gradually increased from larvae to 7 day-old pupa.α1-Bdortub expression levels in 1,2 and 3rd instar were about 0.71,1.16 and 4.01 times of that in 10 day-old pupa,respectively.The expression levels in 1,4,and 7 day-old pupa were about 4.22,13.2 and 18.5 folds of that in 10 day-old pupa,respectively.The results suggest that α1-Bdortub might be closely related to development of B.dorsalis.

关 键 词：橘小实蝇 α微管蛋白 基因克隆 实时荧光定量PCR 

分 类 号：Q965.9[生物学—昆虫学]