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双向蛾卵壳启动子在转基因果蝇中的发育特异性

Developmental specificity of a bidirectional moth chorion promoter in transgenic Drosophila.

作者信息

Fenerjian M G, Kafatos F C

机构信息

Department of Cellular and Developmental Biology, Harvard University, Cambridge, Massachusetts 02138.

出版信息

Dev Biol. 1994 Jan;161(1):37-47. doi: 10.1006/dbio.1994.1005.

Abstract

The developmentally specific activity of the 326-bp promoter that is shared by the paired 18 and 401 chorion genes of the silkmoth Antheraea pernyi was studied in transgenic Drosophila melanogaster. Bidirectional activity of this promoter is demonstrated in a novel dual reporter vector, containing lacZ on one side and CAT on the other. The single chorion-specific hexamer of this promoter, TCACGT, is essential for expression in both orientations; even single-base substitutions (at positions 2 or 4 of the hexamer) suppress expression. With both reporter genes, ovarian expression is greater from the 401 side of the promoter, although the difference between the two sides is greater for the CAT gene. The intact promoter fragment (which includes some 5' untranslated sequences) is expressed with a high degree of tissue specificity, but a central 184-bp fragment derived from it is also active ectopically in parts of the internal male genitalia. Strong spatial as well as temporal regulation is evident with both the intact promoter and its 184-bp subfragment, assayed in either orientation.

摘要

在转基因黑腹果蝇中研究了家蚕柞蚕成对的18和401绒毛膜基因共有的326 bp启动子的发育特异性活性。在一种新型双报告载体中证明了该启动子的双向活性,该载体一侧含有lacZ,另一侧含有CAT。该启动子的单个绒毛膜特异性六聚体TCACGT对于两个方向的表达都是必不可少的;即使是单碱基替换(在六聚体的第2或4位)也会抑制表达。对于两个报告基因,启动子401侧的卵巢表达更高,尽管两个方向之间的差异对于CAT基因更大。完整的启动子片段(包括一些5'非翻译序列)以高度的组织特异性表达,但从中衍生的184 bp中央片段在内雄性生殖器的部分区域也异位活跃。完整启动子及其184 bp亚片段在任一方向上进行检测时,都表现出明显的空间和时间调控。

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