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肝脏5-氨基酮戊酸合酶的分子克隆

Molecular cloning of hepatic 5-aminolevulinate synthase.

作者信息

Borthwick I A, Srivastava G, Hobbs A A, Pirola B A, Brooker J D, May B K, Elliott W H

出版信息

Eur J Biochem. 1984 Oct 1;144(1):95-9. doi: 10.1111/j.1432-1033.1984.tb08435.x.

Abstract

Hepatic 5-aminolevulinate synthase was induced in chick embryos by administration of the porphyrinogenic drugs 2-allyl-2-isopropylacetamide and 3,5-diethoxycarbonyl-1,4-dihydrocollidine. A cDNA library was constructed from drug-induced liver mRNA and clones containing sequences coding for 5-aminolevulinate synthase were identified by hybrid-selected translation. The identity of these clones was confirmed by comparing DNA sequence data with the amino acid sequence of peptides from purified 5-aminolevulinate synthase. From Northern blot analysis the size of the mRNA for 5-aminolevulinate synthase was estimated to be 2800 base pairs, approximately 600 base pairs more than that required to code for the primary translation product of relative molecular mass 74000.

摘要

通过给予致卟啉药物2-烯丙基-2-异丙基乙酰胺和3,5-二乙氧羰基-1,4-二氢可力丁,在鸡胚中诱导出肝脏5-氨基酮戊酸合酶。从药物诱导的肝脏mRNA构建了一个cDNA文库,并通过杂交选择翻译鉴定出含有编码5-氨基酮戊酸合酶序列的克隆。通过将DNA序列数据与来自纯化的5-氨基酮戊酸合酶的肽的氨基酸序列进行比较,证实了这些克隆的身份。通过Northern印迹分析,估计5-氨基酮戊酸合酶的mRNA大小为2800个碱基对,比编码相对分子质量为74000的初级翻译产物所需的碱基对大约多600个。

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