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编码日本萤火虫(Luciola lateralis)荧光素酶的cDNA克隆在大肠杆菌中的分子克隆与表达。

Molecular cloning and expression in Escherichia coli of a cDNA clone encoding luciferase of a firefly, Luciola lateralis.

作者信息

Tatsumi H, Kajiyama N, Nakano E

机构信息

Research and Development Division, Kikkoman Corporation, Noda, Japan.

出版信息

Biochim Biophys Acta. 1992 Jun 15;1131(2):161-5. doi: 10.1016/0167-4781(92)90071-7.

Abstract

We have cloned a cDNA encoding Luciola lateralis (a common firefly in Japan) luciferase from a cDNA library of lantern poly(A)+ RNA, using a cDNA of L. cruciata (another common firefly in Japan) luciferase as a probe. The primary structure of L. lateralis luciferase deduced from the nucleotide sequence was shown to consist of 548 amino acids with a molecular weight of 60,132. Sequence comparison indicates that L. lateralis luciferase has significant sequence identity (94%) to L. cruciata luciferase, and that it has less sequence similarity (67%) to Photinus pyralis (a North American firefly) luciferase. The isolated cDNA clone, when introduced into Escherichia coli, directed the synthesis of enzymatically active luciferase under the control of the lacZ promoter.

摘要

我们使用日本另一种常见萤火虫——姬萤(L. cruciata)荧光素酶的cDNA作为探针,从灯笼多聚腺苷酸加尾(poly(A)+)RNA的cDNA文库中克隆了编码日本扁萤(Luciola lateralis,日本一种常见萤火虫)荧光素酶的cDNA。从核苷酸序列推导的日本扁萤荧光素酶的一级结构显示由548个氨基酸组成,分子量为60,132。序列比较表明,日本扁萤荧光素酶与姬萤荧光素酶具有显著的序列同一性(94%),而与北美萤火虫(Photinus pyralis)荧光素酶的序列相似性较低(67%)。分离得到的cDNA克隆导入大肠杆菌后,在lacZ启动子的控制下指导合成具有酶活性的荧光素酶。

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