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Two Alternatively Spliced Transcripts from the Drosophila period Gene Rescue Rhythms Having Different Molecular and Behavioral Characteristics

机译:果蝇时期的两个交替剪接的转录本,具有不同的分子和行为特征的基因抢救节律

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摘要

The period (per) and timeless (tim) genes encode key components of the circadian oscillator in Drosophila melanogaster. The per gene is thought to encode three transcripts via differential splicing (types A, B, and C) that give rise to three proteins. Since the three per mRNA types were based on the analysis of cDNA clones, we tested whether these mRNA types were present in vivo by RNase protection assays and reverse transcriptase-mediated PCR. The results show that per generates two transcript types that differ only by the presence (type A) or absence (type B′) of an alternative intron in the 3′ untranslated region. Transgenic flies containing transgenes that produce only type B′ transcripts (perB′), type A transcripts (perA), or both transcripts (perG) rescue locomotor activity rhythms with average periods of 24.7, 25.4, and 24.4 h, respectively. Although no appreciable differences in type A and type B′ mRNA cycling were observed, a slower accumulation of PER in flies making only type A transcripts suggests that the intron affects the translation of per mRNA.
机译:周期(每个)和永恒(tim)基因编码果蝇中昼夜节律振荡器的关键成分。每个基因被认为通过产生三个蛋白质的差异剪接(A,B和C型)编码三个转录本。由于每种mRNA的三种类型均基于cDNA克隆的分析,因此我们通过RNase保护试验和逆转录酶介导的PCR检测了这些mRNA类型是否在体内存在。结果表明,每个生成的转录本类型仅在3'非翻译区中存在一个替代内含子(A型)或不存在(B'型)而不同。含有仅产生B'型转录本(perB'),A型转录本(perA)或两种转录本(perG)的转基因果蝇均能挽救运动活动节律,平均周期分别为24.7、25.4和24.4 h。尽管未观察到A型和B'mRNA循环的明显差异,但仅形成A型转录物的果蝇中PER的积累较慢,这表明内含子会影响per mRNA的翻译。

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