家蚕转录因子AP-4基因cDNA的分子克隆与生物信息学分析  

Molecular Cloning and Bioinformatics Analysis of cDNA of Bombyx mori Transcription Factor AP-4

作　　者：李佳梅[1] 赵浩勤[2] 张国政[1,2] 夏定国[1] 张业顺[1] 唐顺明[1] 沈兴家[1] 

机构地区：[1]中国农业科学院蚕业研究所,农业部家蚕生物技术重点开放实验室,江苏镇江212018 [2]江苏科技大学,江苏镇江212003

出　　处：《蚕业科学》2008年第2期211-218,共8页ACTA SERICOLOGICA SINICA

基　　金：国家高技术研究发展计划“863”项目(编号2006AA10A-119);江苏省自然科学基金前期预研重大项目(编号BK2004206)

摘　　要：AP-4(activator protein 4)是一种转录因子,在生物的生长发育中有重要作用。根据家蚕表达序列标签(EST)并利用cDNA末端快速扩增(RACE)方法克隆了家蚕AP-4(Bombyx mori activator protein4)基因,结合生物信息学方法对所获的序列进行开放阅读框、序列同源性分析,预测了AP-4蛋白的理化性质。获得的家蚕AP-4基因cDNA的全长为1621bp,其开放阅读框为996bp,编码331个氨基酸,基因由3个外显子和2个内含子组成。同源性比对表明该基因推导的氨基酸序列与棉铃虫AP-4和谷蛀虫AP-4推测的蛋白同源性分别为76%和54%,第45-99位氨基酸序列是一个典型的保守结构域。实时定量RT-PCR显示该基因在所检测家蚕的各发育时期中,除幼虫1龄和蛹期第4天无表达外,其它时期均有表达,其中幼虫3龄和4龄期的表达量较高。AP-4 is a transcription factor which plays an important role in the progress of growth and development. Based on the information of silkworm ESTs, the cDNA of Bornbyx rnori activator protein 4 gene（BrnAP-4） was cloned using the strategy of rapid amplification of cDNA ends. Concurrently bioinformatic methods were applied to analyze the obtained sequence and its induced amino acid sequence, including ORF, homologous sequences, physical and chemical properties. The results showed that cDNA of BmAP-4 has 1 621 bp and contains an open reading frame encoding a 331 amino acid protein. The BrnAP-4 gene contains 3 exons and 2 introns. The deduced amino acid has 76% identity to Helicoverpa arrnigera transcription factor AP-4, 54% identity to Triboliurn castaneurn activating enhancer-binding protein 4, and the 45 to 99 amino acid is a typical conserved domain. Realtime quantitative RT-PCR showed that the transcript can be detected in all development stages tested except for 1 st instar and 4th day of pupal stage, especially higher in 3rd and 4th instar.

关 键 词：家蚕 转录因子AP-4 CDNA末端快速扩增 基因克隆 生物信息学 

分 类 号：S881[农业科学—特种经济动物饲养]