LdARL-1 组氨酸标签重组蛋白：通过固定化金属亲和扩张床吸附进行纯化

LdARL-1 His-tagged recombinant protein: purification by immobilized metal affinity expanded bed adsorption.

作者信息

Sahin Annelise, Tetaud Emmanuel, Merlin Gilles, Santarelli Xavier

机构信息

CNRS UMR 5162 Génomique Fonctionnelle des Trypanosomatides, Université Victor Segalen Bordeaux 2, 146 rue Léo Saignat, 33076 Bordeaux cedex, France.

出版信息

J Chromatogr B Analyt Technol Biomed Life Sci. 2005 Apr 15;818(1):19-22. doi: 10.1016/j.jchromb.2004.10.022.

DOI:10.1016/j.jchromb.2004.10.022

PMID:15722039

Abstract

Previously we have cloned three ADP-ribosylation factor-like (ARL) genes from the parasitic protozoan Leishmania donovani: LdARL-3A and 3B, LdARL-1. LdARL-3A was previously purified as an active native form, which was able to bind GTP in vitro. In this paper, we have performed the production and the purification of Histidine-tagged (His-tagged) LdARL-1 recombinant protein by immobilized metal affinity chromatography (IMAC) using expanded bed adsorption (EBA) technology. This protein was purified with more than 95% purity and could be successfully used for GTP-binding assay.

摘要

此前，我们已从寄生原生动物杜氏利什曼原虫中克隆出三个ADP-核糖基化因子样（ARL）基因：LdARL-3A和3B、LdARL-1。LdARL-3A先前已被纯化出活性天然形式，它能够在体外结合GTP。在本文中，我们利用扩张床吸附（EBA）技术，通过固定化金属亲和层析（IMAC）法进行了组氨酸标签（His标签）LdARL-1重组蛋白的制备和纯化。该蛋白的纯度超过95%，可成功用于GTP结合测定。