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黑腹果蝇转座子介导的抑制因子——黑貂抑制因子的分子克隆

Molecular cloning of suppressor of sable, a Drosophila melanogaster transposon-mediated suppressor.

作者信息

Chang D Y, Wisely B, Huang S M, Voelker R A

出版信息

Mol Cell Biol. 1986 May;6(5):1520-8. doi: 10.1128/mcb.6.5.1520-1528.1986.

Abstract

A hybrid dysgenesis-induced allele [su(s)w20] associated with a P-element insertion was used to clone sequences from the su(s) region of Drosophila melanogaster by means of the transposon-tagging technique. Cloned sequences were used to probe restriction enzyme-digested DNAs from 22 other su(s) mutations. None of three X-ray-induced or six ethyl methanesulfonate-induced su(s) mutations possessed detectable variation. Seven spontaneous, four hybrid dysgenesis-induced, and two DNA transformation-induced mutations were associated with insertions within 2.0 kilobases (kb) of the su(s)w20 P-element insertion site. When the region of DNA that included the mutational insertions was used to probe poly(A)+ RNAs, a 5-kb message was detected in wild-type RNA that was present in greatly reduced amounts in two su(s) mutations. By using strand-specific probes, the direction of transcription of the 5-kb message was determined. The mutational insertions lie in DNA sequences near the 5' end of the 5-kb message. Three of the seven spontaneous su(s) mutations are associated with gypsy insertions, but they are not suppressible by su(Hw).

摘要

利用与P因子插入相关的杂种发育不全诱导等位基因[su(s)w20],通过转座子标签技术从黑腹果蝇的su(s)区域克隆序列。克隆的序列用于探测来自其他22个su(s)突变的经限制性内切酶消化的DNA。三个X射线诱导的或六个甲磺酸乙酯诱导的su(s)突变均未检测到可察觉的变异。七个自发突变、四个杂种发育不全诱导突变和两个DNA转化诱导突变与su(s)w20 P因子插入位点2.0千碱基(kb)范围内的插入有关。当包含突变插入的DNA区域用于探测聚腺苷酸加尾RNA(poly(A)+ RNAs)时,在野生型RNA中检测到一个5-kb的信使RNA,而在两个su(s)突变中其含量大幅减少。通过使用链特异性探针,确定了5-kb信使RNA的转录方向。突变插入位于5-kb信使RNA 5'端附近的DNA序列中。七个自发的su(s)突变中有三个与gypsy插入有关,但它们不能被su(Hw)抑制。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/82e1/367677/1970962a51e9/molcellb00089-0173-a.jpg

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