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通过克隆互补DNA确定的卵类粘蛋白信使核糖核酸的一级序列。

Primary sequence of ovomucoid messenger RNA as determined from cloned complementary DNA.

作者信息

Catterall J F, Stein J P, Kristo P, Means A R, O'Malley B W

出版信息

J Cell Biol. 1980 Nov;87(2 Pt 1):480-7. doi: 10.1083/jcb.87.2.480.

Abstract

Ovomucoid messenger RNA (mRNAom) comprises approximately 8% of the total mRNA in the estrogen-stimulated oviduct. The recombinant plasmid pOM100 contained DNA complementary to the 3' end of mRNAom. DNA complementary to the 5' end of mRNAom was obtained from a partially purified preparation of mRNAom by polymerization by reverse transcriptase in the presence of a restriction fragment primer from pOM100. The complementary DNA mixture was amplified by molecular cloning using poly dG/dC tailing to form recombinant bacterial plasmids. Recombinant plasmids containing ovomucoid DNA sequences were selected by in situ hybridization to 32P-labeled pOM100 fragments. The longest plasmid containing ovomucoid DNA sequences was designated pOM502. The complete DNA sequence of both pOM100 and pOM502 was determined. The two plasmids appear to contain sequences complementary to the entire length of mRNAom. The nucleic acid sequence agrees with the known amino acid sequences for both ovomucoid and its N-terminal signal peptide. Highly homologous sequences occur in two regions that coincide with structural domains of the protein. Comparison of the sequence of mRNAom with that for other eucaryotic mRNAs allowed identification of possible functional regions in the mRNA molecule.

摘要

卵类粘蛋白信使核糖核酸(mRNAom)约占雌激素刺激的输卵管中总信使核糖核酸的8%。重组质粒pOM100含有与mRNAom 3'末端互补的DNA。通过在来自pOM100的限制性片段引物存在下用逆转录酶进行聚合反应,从部分纯化的mRNAom制剂中获得了与mRNAom 5'末端互补的DNA。通过使用聚dG/dC加尾进行分子克隆来扩增互补DNA混合物,以形成重组细菌质粒。通过与32P标记的pOM100片段进行原位杂交来选择含有卵类粘蛋白DNA序列的重组质粒。含有卵类粘蛋白DNA序列的最长质粒被命名为pOM502。测定了pOM100和pOM502的完整DNA序列。这两个质粒似乎包含与mRNAom全长互补的序列。核酸序列与卵类粘蛋白及其N端信号肽的已知氨基酸序列一致。高度同源的序列出现在与该蛋白质结构域一致的两个区域。将mRNAom的序列与其他真核信使核糖核酸的序列进行比较,使得能够鉴定出mRNA分子中可能的功能区域。

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Isolation and characterization of the chicken ovomucoid gene.鸡卵类粘蛋白基因的分离与鉴定
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本文引用的文献

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A membrane-filter technique for the detection of complementary DNA.一种用于检测互补DNA的膜过滤技术。
Biochem Biophys Res Commun. 1966 Jun 13;23(5):641-6. doi: 10.1016/0006-291x(66)90447-5.

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