一种通过基因产物活性来鉴定基因的生化基因组学方法。

A biochemical genomics approach for identifying genes by the activity of their products.

作者信息

Martzen M R, McCraith S M, Spinelli S L, Torres F M, Fields S, Grayhack E J, Phizicky E M

机构信息

Department of Biochemistry and Biophysics, University of Rochester School of Medicine, Rochester, NY 14642, USA.

出版信息

Science. 1999 Nov 5;286(5442):1153-5. doi: 10.1126/science.286.5442.1153.

DOI:10.1126/science.286.5442.1153

PMID:10550052

Abstract

For the identification of yeast genes specifying biochemical activities, a genomic strategy that is rapid, sensitive, and widely applicable was developed with an array of 6144 individual yeast strains, each containing a different yeast open reading frame (ORF) fused to glutathione S-transferase (GST). For the identification of ORF-associated activities, strains were grown in defined pools, and GST-ORFs were purified. Then, pools were assayed for activities, and active pools were deconvoluted to identify the source strains. Three previously unknown ORF-associated activities were identified with this strategy: a cyclic phosphodiesterase that acts on adenosine diphosphate-ribose 1"-2" cyclic phosphate (Appr>p), an Appr-1"-p-processing activity, and a cytochrome c methyltransferase.

摘要

为了鉴定决定生化活性的酵母基因，开发了一种快速、灵敏且广泛适用的基因组策略，该策略使用了一组6144个单独的酵母菌株，每个菌株都包含一个与谷胱甘肽S-转移酶（GST）融合的不同酵母开放阅读框（ORF）。为了鉴定与ORF相关的活性，将菌株在特定的混合培养物中培养，然后纯化GST-ORF。接着，对混合培养物进行活性测定，并对有活性的混合培养物进行反卷积分析以鉴定来源菌株。通过该策略鉴定出了三种以前未知的与ORF相关的活性：一种作用于二磷酸腺苷核糖1"-2"环磷酸酯（Appr>p）的环磷酸二酯酶、一种Appr-1"-p加工活性以及一种细胞色素c甲基转移酶。