乳草长蝽Ubx基因克隆及多转录本分析  

Cloning and multiple transcript analysis of Ubx in the large milkweed bug,Oncopeltus fasciatus(Hemiptera:Lygaeidae)

作　　者：田晓轩[1] 谢强[1] 卜文俊[1] 

机构地区：[1]南开大学生命科学学院昆虫学研究所,天津300071

出　　处：《昆虫学报》2011年第4期390-396,共7页Acta Entomologica Sinica

基　　金：国家自然科学基金项目(30725005;J0930005)

摘　　要：针对非完全变态类昆虫发育关键基因的研究相对匮乏,尤其缺少Hox基因家族的基因结构和序列信息。为了研究Hox基因家族成员之一的Ubx基因在非完全变态类昆虫中的结构特点,本实验选取乳草长蝽Oncopeltus fasciatus(Dallas,1852)为代表,应用RACE和RT-PCR技术,对其Ubx基因的全长开放阅读框进行克隆。结果显示:乳草长蝽Ubx基因(Of-Ubx)开放阅读框全长888bp,推测的完整蛋白含有295个氨基酸。Southern blot证实Ubx基因以单拷贝形式存在且含有内含子。在Of-Ubx的YPWM基序和同源异型结构域之间存在选择性剪接位点,可产生3种不同转录本。分析以上实验结果,发现乳草长蝽与黑腹果蝇Drosophila melanogaster(Meigen,1830)的Ubx基因拥有相似的剪接位置、剪接体组合和边界序列,提示它们很可能具有相似的剪接机理。这是Ubx基因的多转录本现象在昆虫纲中果蝇属以外类群中的首次详尽报道。The researches focusing on key developmental genes in non-holometabola insects are relatively insufficient,especially for the reports on the structure and sequences of Hox genes. In order to understand the structure of Ubx gene （a member of Hox genes） in non-holometabola insects,we selected the large milkweed bug,Oncopeltus fasciatus （Dallas,1852）,as a representative,and used RACE and RT-PCR to clone the whole ORF of Ubx gene. The results showed that the full-length ORF of Ubx gene of O. fasciatus （Of-Ubx） is 888 bp,encoding 295 aa. Southern blot analysis verified that Of-Ubx exists as a single copy and has introns within it. We found alternative splicing sites between YPWM and homeodomain motif,which lead to 3 types of transcript variants. Compared with the Ubx gene from Drosophila melanogaster （Meigen,1830）,Of-Ubx had similar splicing sites,conserved elements around splicing sites and isoform combination,suggesting that their splicing mechanism should be similar. This is the first detailed report about the multiple transcripts of Ubx genes within Insecta except for the genus Drosophila.

关 键 词：乳草长蝽 Ubx基因 克隆 开放阅读框 多转录本 

分 类 号：Q966[生物学—昆虫学]