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正常、突变和截短的小鼠α-珠蛋白基因的体外转录

In vitro transcription of normal, mutant, and truncated mouse alpha-globin genes.

作者信息

Talkington C A, Nishioka Y, Leder P

出版信息

Proc Natl Acad Sci U S A. 1980 Dec;77(12):7132-6. doi: 10.1073/pnas.77.12.7132.

DOI:10.1073/pnas.77.12.7132
PMID:6938959
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC350455/
Abstract

We have transcribed various alpha-globin gene-containing templates by using a simplified in vitro system and have shown that the 5' end of each of the RNA polymerase II-specific transcripts corresponds to that of authentic alpha-globin mRNA. Using a truncated alpha-globin gene fragment, we then localized the promotor region of the adult alpha-globin gene to a 138-base-pair fragment of alpha-globin DNA, 141 nucleotides of which precede the initiation site. In contrast, when two naturally occurring alpha-globin gene mutants were tested as templates, no RNA polymerase II-dependent initiation was detected. Comparison of the nucleotide sequences of the normal and mutant alpha-globin genes has allowed us to focus upon nucleotide positions that might be essential for promoter activity.

摘要

我们使用一种简化的体外系统转录了各种含α-珠蛋白基因的模板，并表明每个RNA聚合酶II特异性转录本的5'端与真实α-珠蛋白mRNA的5'端相对应。然后，我们使用一个截短的α-珠蛋白基因片段，将成人α-珠蛋白基因的启动子区域定位到α-珠蛋白DNA的一个138个碱基对的片段上，其中141个核苷酸位于起始位点之前。相比之下，当测试两个天然存在的α-珠蛋白基因突变体作为模板时，未检测到RNA聚合酶II依赖性起始。对正常和突变α-珠蛋白基因的核苷酸序列进行比较，使我们能够关注可能对启动子活性至关重要的核苷酸位置。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9ced/350455/75495693ea85/pnas00499-0197-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9ced/350455/6305e918377f/pnas00499-0195-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9ced/350455/6dfa3689cb7a/pnas00499-0196-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9ced/350455/b18f272b89cd/pnas00499-0197-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9ced/350455/75495693ea85/pnas00499-0197-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9ced/350455/6305e918377f/pnas00499-0195-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9ced/350455/6dfa3689cb7a/pnas00499-0196-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9ced/350455/b18f272b89cd/pnas00499-0197-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9ced/350455/75495693ea85/pnas00499-0197-b.jpg

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本文引用的文献

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Cell. 1980 Jul;20(3):691-9. doi: 10.1016/0092-8674(80)90315-3.
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Mouse globin system: a functional and evolutionary analysis.小鼠珠蛋白系统：功能与进化分析
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Specific in vitro initiation of transcription on conalbumin and ovalbumin genes and comparison with adenovirus-2 early and late genes.
烟草中一种短散在重复元件的分子特征分析，该元件与特定tRNA具有序列同源性。
Proc Natl Acad Sci U S A. 1993 Jul 15;90(14):6562-6. doi: 10.1073/pnas.90.14.6562.
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Simian virus 40 major late promoter: an upstream DNA sequence required for efficient in vitro transcription.猴病毒40主要晚期启动子：体外高效转录所需的上游DNA序列。
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Definition of the ovalbumin gene promoter by transfer of an ovalglobin fusion gene into cultured cells.通过将卵清球蛋白融合基因转入培养细胞来定义卵清蛋白基因启动子。
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