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Notch的分子克隆,Notch是一个影响黑腹果蝇神经发生的基因座。

Molecular cloning of Notch, a locus affecting neurogenesis in Drosophila melanogaster.

作者信息

Artavanis-Tsakonas S, Muskavitch M A, Yedvobnick B

出版信息

Proc Natl Acad Sci U S A. 1983 Apr;80(7):1977-81. doi: 10.1073/pnas.80.7.1977.

Abstract

The Notch locus is one of the best characterized loci in Drosophila melanogaster in terms of its genetic structure and developmental effects. Mutations in this locus profoundly affect the differentiation of the early embryo. Using an inversion involving the Notch locus and previously cloned sequences, we have isolated chromosomal segments from the Notch region (3C7) encompassing 80 kilobases (kb) of DNA. Based on comparison between mutant and wild-type DNA, we have positioned cloned sequences within the Notch genetic map; furthermore, we have defined a region of approximately 40 kb within which the structural lesions correlating with all Notch alleles mapped to date appear to reside. We have examined the transcriptional activity of the cloned sequences during ontogeny and find a single size class of poly(A)+ RNA, 10.5 kb long, that is homologous to sequences within this 40-kb region. We conclude that DNA sequences belonging to the Notch locus have been cloned and that the 10.5-kb poly(A)+ RNA is essential for wild-type Notch function. We discuss these structural and transcriptional data in light of the existing genetic and developmental characterization of the Notch locus.

摘要

就其遗传结构和发育效应而言,Notch基因座是黑腹果蝇中特征最明确的基因座之一。该基因座的突变会深刻影响早期胚胎的分化。利用一个涉及Notch基因座和先前克隆序列的倒位,我们从Notch区域(3C7)分离出了包含80千碱基(kb)DNA的染色体片段。基于突变型和野生型DNA之间的比较,我们已将克隆序列定位在Notch遗传图谱中;此外,我们还确定了一个约40 kb的区域,迄今为止定位的所有Notch等位基因相关的结构损伤似乎都位于该区域内。我们检查了克隆序列在个体发育过程中的转录活性,发现了一类单一大小的多聚腺苷酸(poly(A))+ RNA,长度为10.5 kb,与该40 kb区域内的序列同源。我们得出结论,属于Notch基因座的DNA序列已被克隆,并且10.5 kb的多聚腺苷酸(poly(A))+ RNA对野生型Notch功能至关重要。我们根据Notch基因座现有的遗传和发育特征来讨论这些结构和转录数据。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6431/393735/7952af7f9d2f/pnas00633-0199-a.jpg

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