古细菌中的逆转录酶。嗜盐嗜盐菌中一种引发酶-逆转录酶复合物的纯化与特性分析。

Reverse transcriptase in archaebacteria. Purification and characterization of a primase-reverse-transcriptase complex from Halobacterium halobium.

作者信息

Ben-Mahrez K, Sorokine I, Nakayama M, Kohiyama M

机构信息

Institut Jacques Monod, Centre National de la Recherche Scientifique, Université Paris, France.

出版信息

Eur J Biochem. 1991 Jan 1;195(1):157-62. doi: 10.1111/j.1432-1033.1991.tb15689.x.

DOI:10.1111/j.1432-1033.1991.tb15689.x

PMID:1703956

Abstract

A primase-reverse-transcriptase of Halobacterium halobium was purified by column chromatography on DEAE-cellulose, hydroxyapatite and carboxymethyl-cellulose, followed by sedimentation on a glycerol gradient. The enzyme is a multifunctional enzyme containing reverse transcriptase. DNA polymerase and RNase H activities and does not require a performed primer to initiate DNA synthesis. Using a single-stranded DNA as template, this enzyme synthesizes oligonucleotides (8-12 bases) that can be used a primer by Escherichia coli DNA nucleotidyltransferase I (DNA polymerase I, Klenow fragment). Two polypeptides of 67 and 57 kDa were found after 14750-fold purification of the enzyme.

摘要

嗜盐栖热菌的一种引发酶逆转录酶通过在二乙氨基乙基纤维素、羟基磷灰石和羧甲基纤维素上进行柱层析，随后在甘油梯度上进行沉降而被纯化。该酶是一种含有逆转录酶、DNA聚合酶和核糖核酸酶H活性的多功能酶，并且不需要预先存在的引物来起始DNA合成。以单链DNA作为模板，这种酶合成的寡核苷酸（8 - 12个碱基）可被大肠杆菌DNA核苷酸转移酶I（DNA聚合酶I，克列诺片段）用作引物。在该酶经过14750倍纯化后，发现了两条分别为67 kDa和57 kDa的多肽。