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拟南芥乙醇脱氢酶基因的分子克隆与DNA序列

Molecular cloning and DNA sequence of the Arabidopsis thaliana alcohol dehydrogenase gene.

作者信息

Chang C, Meyerowitz E M

出版信息

Proc Natl Acad Sci U S A. 1986 Mar;83(5):1408-12. doi: 10.1073/pnas.83.5.1408.

Abstract

Arabidopsis thaliana provides an excellent experimental plant system for molecular genetics because of its remarkably small genome size, near absence of dispersed middle repetitive DNA, and short life cycle. We have cloned and determined the nucleotide sequence of a single-copy gene from A. thaliana likely to be the gene encoding alcohol dehydrogenase (ADH; alcohol:NAD+ oxidoreductase, EC 1.1.1.1). The gene was isolated from a random recombinant library by cross-hybridization with a maize Adh1 gene probe. The DNA sequence contains an open reading frame capable of encoding a polypeptide the same length as maize ADH1 and ADH2 (379 amino acids) and having approximately equal to 80% homology with both maize enzymes. This open reading frame is interrupted by six introns whose positions are conserved with six of the nine intron positions present in both maize genes. The 5' and 3' untranslated regions are, respectively, 58 and 204 base pairs long. Sequences important for eukaryotic gene expression such as the TATA box, polyadenylylation signal, and intron splicesite sequences are found in the expected locations. The gene hybridizes to a specific anaerobically induced RNA in Arabidopsis whose appearance correlates with the anaerobic induction of Arabidopsis ADH protein.

摘要

由于拟南芥基因组显著小、几乎不存在分散的中度重复DNA且生命周期短,它为分子遗传学提供了一个出色的实验植物系统。我们已克隆并确定了拟南芥中一个单拷贝基因的核苷酸序列,该基因可能是编码乙醇脱氢酶(ADH;乙醇:NAD⁺氧化还原酶,EC 1.1.1.1)的基因。通过与玉米Adh1基因探针交叉杂交,从随机重组文库中分离出该基因。DNA序列包含一个开放阅读框,能够编码与玉米ADH1和ADH2长度相同(379个氨基酸)且与这两种玉米酶具有约80%同源性的多肽。这个开放阅读框被六个内含子打断,其位置与两个玉米基因中九个内含子位置中的六个保守。5'和3'非翻译区分别长58和204个碱基对。在预期位置发现了对真核基因表达重要的序列,如TATA框、多聚腺苷酸化信号和内含子剪接位点序列。该基因与拟南芥中一种特定的厌氧诱导RNA杂交,其出现与拟南芥ADH蛋白的厌氧诱导相关。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ba01/323085/ba137fc4162f/pnas00309-0248-a.jpg

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