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嗜热栖热菌DNA连接酶(大肠杆菌DNA连接酶的同源物)的克隆、核苷酸序列及工程化表达

Cloning, nucleotide sequence, and engineered expression of Thermus thermophilus DNA ligase, a homolog of Escherichia coli DNA ligase.

作者信息

Lauer G, Rudd E A, McKay D L, Ally A, Ally D, Backman K C

机构信息

BioTechnica Diagnostics, Inc., Cambridge, Massachusetts 02138.

出版信息

J Bacteriol. 1991 Aug;173(16):5047-53. doi: 10.1128/jb.173.16.5047-5053.1991.

Abstract

We have cloned and sequenced the gene for DNA ligase from Thermus thermophilus. A comparison of this sequence and those of other ligases reveals significant homology only with that of Escherichia coli. The overall amino acid composition of the thermophilic ligase and the pattern of amino acid substitutions between the two proteins are consistent with compositional biases in other thermophilic enzymes. We have engineered the expression of the T. thermophilus gene in Escherichia coli, and we show that E. coli proteins may be substantially removed from the thermostable ligase by a simple heat precipitation step.

摘要

我们已经克隆并测序了嗜热栖热菌DNA连接酶的基因。将该序列与其他连接酶的序列进行比较后发现,只有与大肠杆菌的连接酶序列有显著同源性。嗜热连接酶的整体氨基酸组成以及这两种蛋白质之间的氨基酸取代模式,与其他嗜热酶的组成偏好一致。我们已经设计了嗜热栖热菌基因在大肠杆菌中的表达,并且表明通过简单的热沉淀步骤,大肠杆菌蛋白可从耐热连接酶中大量去除。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b292/208194/ec6a6a7a23aa/jbacter00106-0148-a.jpg

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