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人源蛋白pLG72在大肠杆菌中的表达及体外重折叠

Expression in Escherichia coli and in vitro refolding of the human protein pLG72.

作者信息

Molla Gianluca, Bernasconi Mariagrazia, Sacchi Silvia, Pilone Mirella S, Pollegioni Loredano

机构信息

Department of Biotechnology and Molecular Sciences, University of Insubria, Via J. H. Dunant, 3, 21100 Varese, Italy.

出版信息

Protein Expr Purif. 2006 Mar;46(1):150-5. doi: 10.1016/j.pep.2005.08.003. Epub 2005 Sep 7.

Abstract

Recently, genes coding for pLG72 and d-amino acid oxidase have been related to schizophrenia, a widespread psychiatric disorder that affects about 1% of population. pLG72 is a puzzling, novel protein present only in primates and proposed to be an activator of d-amino acid oxidase. Here we report on the overexpression of wild-type and His-tagged pLG72 in Escherichia coli. Both variants form inclusion bodies and have been refolded and purified to homogeneity: the acquisition of secondary and tertiary structure was demonstrated by CD spectroscopy. A figure of approximately 70 mg of pure protein per liter of fermentation broth was achieved.

摘要

最近,编码pLG72和D-氨基酸氧化酶的基因已被证实与精神分裂症有关,精神分裂症是一种广泛存在的精神疾病,影响着约1%的人口。pLG72是一种仅存在于灵长类动物中的令人费解的新型蛋白质,被认为是D-氨基酸氧化酶的激活剂。在此,我们报道了野生型和His标签化的pLG72在大肠杆菌中的过表达。这两种变体均形成包涵体,并已复性和纯化至均一性:通过圆二色光谱证明了二级和三级结构的获得。每升发酵液可获得约70毫克纯蛋白。

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