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真核细胞中自主复制序列(ARS)元件的鉴定。

Identification of autonomously replicating sequence (ARS) elements in eukaryotic cells.

作者信息

Clyne R K, Kelly T J

机构信息

Department of Molecular Biology and Genetics, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205, USA.

出版信息

Methods. 1997 Nov;13(3):221-33. doi: 10.1006/meth.1997.0522.

Abstract

Autonomously replicating sequence (ARS) elements were first identified in the budding yeast Saccharomyces cerevisiae as chromosomal DNA fragments that promoted high frequency of transformation and extrachromosomal maintenance of plasmid DNA. These specific sequence elements were subsequently shown to function as origins of DNA replication. Detailed analysis of the structure and function of ARS elements has been limited largely to S. cerevisiae and more recently the fission yeast Schizosaccharomyces pombe. Characterization of ARS activity in other eukaryotes is far less complete. Here we describe the ARS assay developed in yeast and its application to the study of origin function in other eukaryotes. Other available methods for detecting autonomous replication in these systems are also presented.

摘要

自主复制序列(ARS)元件最初是在芽殖酵母酿酒酵母中作为染色体DNA片段被鉴定出来的,这些片段可促进质粒DNA的高频转化和染色体外维持。随后发现这些特定的序列元件可作为DNA复制的起点。ARS元件的结构和功能的详细分析主要限于酿酒酵母,最近也扩展到了裂殖酵母粟酒裂殖酵母。在其他真核生物中对ARS活性的表征则远没有那么完整。在这里,我们描述了在酵母中开发的ARS检测方法及其在研究其他真核生物中起点功能的应用。还介绍了在这些系统中检测自主复制的其他可用方法。

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