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通过P因子介导的转化分析黑腹果蝇Sgs-4基因的调控序列。

Regulatory sequences of the Sgs-4 gene of Drosophila melanogaster analysed by P element-mediated transformation.

作者信息

Hofmann A, Keinhorst A, Krumm A, Korge G

机构信息

Institut für Genetik, Freie Universität Berlin.

出版信息

Chromosoma. 1987;96(1):8-17. doi: 10.1007/BF00285877.

Abstract

The X chromosomally located allele Sgs-4c for a larval secretion protein of Drosophila melanogaster is normally expressed in female larvae of the strain Oregon R and is hyperexpressed in male larvae exhibiting dosage compensation; the allele Sgs-4d in the strain Samarkand is weakly expressed and is not hyperexpressed in male larvae showing a dosage effect. P element-mediated transformation of upstream DNA sequences from both alleles combined with Sgs-4d coding and downstream sequences was performed to localize sequences which are responsible for the level of gene expression and for hyperexpression of Sgs-4c in male larvae. Our results demonstrate that weak expression and dosage effect are inherited with the upstream region from -1 to -838. This Samarkand fragment differs from the homologous Oregon R region only by a C to T transition at -344 which lies within an assumed binding sequence for the ecdysone receptor complex of dyad base symmetry. Replacing the Samarkand upstream region from -1 to -838 by the Oregon R region restores normal Sgs-4 expression and dosage compensation. Hyperexpression in male larvae displays high sensitivity to position effect and is nearly completely inhibited in one transformed line under heterozygous conditions. The integration of an Sgs-4d transposon into a weak spot of polytene chromosome 2L results in a decrease in gene expression. The GTT- and GT-rich regions at -1.2 and -2.0 kb do not obviously influence Sgs-4 expression but possibly play a role in induction of stage-specific chromosome puffing.

摘要

位于X染色体上的黑腹果蝇幼虫分泌蛋白的等位基因Sgs - 4c,在俄勒冈R品系的雌性幼虫中正常表达,在表现出剂量补偿的雄性幼虫中过度表达;撒马尔罕品系中的等位基因Sgs - 4d表达较弱,在表现出剂量效应的雄性幼虫中不发生过度表达。对来自两个等位基因的上游DNA序列与Sgs - 4d编码区及下游序列进行P因子介导的转化,以定位负责基因表达水平以及雄性幼虫中Sgs - 4c过度表达的序列。我们的结果表明,弱表达和剂量效应是由 - 1至 - 838的上游区域遗传而来的。这个撒马尔罕片段与同源的俄勒冈R区域的差异仅在于 - 344处的一个C到T的转换,该位置位于假定的蜕皮激素受体复合物二元碱基对称结合序列内。用俄勒冈R区域替换撒马尔罕 - 1至 - 838的上游区域可恢复正常的Sgs - 4表达和剂量补偿。雄性幼虫中的过度表达对位置效应高度敏感,在杂合条件下的一个转化系中几乎完全受到抑制。将Sgs - 4d转座子整合到多线染色体2L的一个弱点会导致基因表达下降。 - 1.2 kb和 - 2.0 kb处富含GTT和GT的区域对Sgs - 4表达没有明显影响,但可能在阶段特异性染色体胀泡的诱导中起作用。

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