Borthwick I A, Srivastava G, Hobbs A A, Pirola B A, Brooker J D, May B K, Elliott W H
Eur J Biochem. 1984 Oct 1;144(1):95-9. doi: 10.1111/j.1432-1033.1984.tb08435.x.
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.
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。