Diapause-specific gene expression in pupae of the flesh fly Sarcophaga crassipalpis
Diapause-specific gene expression in pupae of the flesh fly Sarcophaga crassipalpis
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DOI：
10.1073/pnas.95.10.5616
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发表时间：
1998-05-12
影响因子：
11.1
通讯作者：
Denlinger, DL
中科院分区：
文献类型：
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作者：
Flannagan, RD;Tammariello, SP;Denlinger, DL
从肉蝇（Sarcophaga crassipalpis）滞育蛹的大脑中分离出的一些cDNA显示出滞育特有的表达模式，为了分离出这些cDNA，采用消除杂交技术筛选了一个滞育蛹的大脑cDNA文库，并选择了与非滞育蛹RNA构建的cDNA探针不杂交的cDNA进行进一步筛选。选择在初始文库筛选中未杂交的95个克隆进行进一步鉴定。然后对这些无性系进行滞育和非滞育蛹多聚(A)(+) Northern印迹检测。二级筛选鉴定出4个滞育上调克隆，7个滞育下调克隆，8个在滞育和非滞育中均表达，75个无表达。通过部分DIVA测序和GenBank的身份搜索，进一步对滞育上调和下调克隆进行了鉴定。我们克隆的cdna与其他基因之间的特性包括与细胞周期进程、应激反应和DNA修复过程相关的基因。结果表明，昆虫滞育不仅仅是基因表达的关闭，而是一种独特的发育途径，其特征是一组新的基因的表达。
Several cDNAs isolated from brains of diapausing pupae of the flesh fly, Sarcophaga crassipalpis, show expression patterns unique to diapause, To isolate such cDNAs a diapause pupal brain cDNA library was screened by using an elimination hybridization technique, and cDNAs that did not hybridize with cDNA probes constructed from the RNA of nondiapausing pupae were selected for further screening. The 95 clones that did not hybridize in the initial library screen were selected for further characterization. These clones were then screened against diapause and nondiapause pupal poly(A)(+) Northern blots. The secondary screen identified 4 diapause-up-regulated clones, 7 diapause-down-regulated clones, 8 clones expressed equally in both diapause and nondiapause, and 75 clones without detectable expression. The diapause-up-regulated and down-regulated clones were further characterized by partial DIVA sequencing and identity searches by using GenBank. Identities between our cloned cDNAs and other genes included those linked to cell cycle progression, stress responses, and DNA repair processes. The results suggest that insect diapause is not merely a shutdown of gene expression but is a unique, developmental pathway characterized by the expression of a novel set of genes.