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人下颌下腺富含脯氨酸蛋白mRNA的无细胞翻译

Cell-free translation of proline-rich protein mRNAS from human submandibular gland.

作者信息

Troxler R F, Belford H S, Oppenheim F G

出版信息

Biochem Biophys Res Commun. 1983 Feb 28;111(1):239-46. doi: 10.1016/s0006-291x(83)80142-9.

Abstract

RNA was isolated from a human submandibular gland and separated into poly A-enriched and poly A-deficient fractions by chromatography on oligo (dT) cellulose. Both of these RNA fractions stimulated methionine incorporation into polypeptides in a reticulocyte lysate translation system. Two in vitro translation products templated by poly A-enriched mRNA were isolated by immunoprecipitation with immune serum directed against human salivary anionic proline-rich protein I. These polypeptides were shown to be precursors of proline-rich proteins on the basis of Mr, affinity for the antiserum, and preferential incorporation of proline. This study is the first to demonstrate cell-free translation of the mRNAs for human proline-rich salivary protein precursors.

摘要

从人的下颌下腺中分离出RNA,并通过在寡聚(dT)纤维素上进行层析,将其分为富含多聚腺苷酸(poly A)和缺乏多聚腺苷酸的组分。这两种RNA组分都能在网织红细胞裂解物翻译系统中刺激蛋氨酸掺入多肽。通过用针对人唾液富含阴离子脯氨酸的蛋白质I的免疫血清进行免疫沉淀,分离出了由富含多聚腺苷酸的mRNA作为模板的两种体外翻译产物。基于分子量、对抗血清的亲和力以及脯氨酸的优先掺入,这些多肽被证明是富含脯氨酸蛋白质的前体。这项研究首次证明了人唾液富含脯氨酸蛋白质前体的mRNA的无细胞翻译。

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