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果蝇多巴脱羧酶基因上游增强异源基因表达的序列基序的鉴定。

Identification of a sequence motif upstream of the Drosophila dopa decarboxylase gene that enhances heterologous gene expression.

作者信息

Hodgetts R B, Patel M S, Piorecky J, Swan A D, Spencer C A

机构信息

Department of Genetics, University of Alberta, Edmonton, Canada.

出版信息

Genome. 1994 Aug;37(4):526-34. doi: 10.1139/g94-075.

Abstract

In this paper we have examined the role that element S, a DNA sequence motif found approximately 215 bp upstream of the Dopa decarboxylase (Ddc) gene, might play in regulating Ddc expression. Nearly identical versions of the element are present upstream of four other Drosophila genes. For two of these, the element appears to be an important component of the upstream regulatory region, since mutations in it reduce expression of the downstream gene. Because an element S polymorphism differentiates the Ddc+ allele of an inbred laboratory strain from the Ddc+4 allele present in a strain isolated from the wild, we decided to test the activity of both forms. Oligonucleotides containing Ddc+ or Ddc+4 versions of element S were synthesized and their ability to drive the expression of an heterologous (Adh) reporter gene at the second molt was examined. Transgenic larvae carrying the element S-Adh fusion constructs consistently exhibited Adh levels that were elevated 1.5-fold above those seen in control organisms. We have also determined the effects of element S in white prepupae and once again, ADH expression levels were significantly above controls in both groups of transformants carrying the element S construct. The results point to a functional role for element S. Since reporter gene expression in third instar larvae was restricted to tissues where ADH is normally found, we conclude that element S is not involved in directing the tissue specificity of Ddc expression.(ABSTRACT TRUNCATED AT 250 WORDS)

摘要

在本文中,我们研究了元素S(一种在多巴脱羧酶(Ddc)基因上游约215 bp处发现的DNA序列基序)在调节Ddc表达中可能发挥的作用。该元素的几乎相同版本存在于其他四个果蝇基因的上游。对于其中两个基因,该元素似乎是上游调控区域的重要组成部分,因为其中的突变会降低下游基因的表达。由于元素S多态性将近交实验室品系的Ddc +等位基因与从野生环境中分离出的品系中存在的Ddc +4等位基因区分开来,我们决定测试这两种形式的活性。合成了含有元素S的Ddc +或Ddc +4版本的寡核苷酸,并检测了它们在第二次蜕皮时驱动异源(Adh)报告基因表达的能力。携带元素S-Adh融合构建体的转基因幼虫始终表现出Adh水平比对照生物体中观察到的水平高出1.5倍。我们还确定了元素S在白色预蛹中的作用,并且再次发现,在携带元素S构建体的两组转化体中,ADH表达水平均明显高于对照。结果表明元素S具有功能作用。由于报告基因在三龄幼虫中的表达仅限于通常发现ADH的组织,我们得出结论,元素S不参与指导Ddc表达的组织特异性。(摘要截短至250字)

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