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不同基因富集方法在鉴定和测序玉米基因空间方面的效用。

Utility of different gene enrichment approaches toward identifying and sequencing the maize gene space.

作者信息

Springer Nathan Michael, Xu Xiequn, Barbazuk W Brad

机构信息

Center for Plant and Microbial Genomics, Department of Plant Biology, University of Minnesota, St. Paul, Minnesota 55108, USA.

出版信息

Plant Physiol. 2004 Oct;136(2):3023-33. doi: 10.1104/pp.104.043323. Epub 2004 Aug 6.

Abstract

Maize (Zea mays) possesses a large, highly repetitive genome, and subsequently a number of reduced-representation sequencing approaches have been used to try and enrich for gene space while eluding difficulties associated with repetitive DNA. This article documents the ability of publicly available maize expressed sequence tag and Genome Survey Sequences (GSSs; many of which were isolated through the use of reduced representation techniques) to recognize and provide coverage of 78 maize full-length cDNAs (FLCs). All 78 FLCs in the dataset were identified by at least three GSSs, indicating that the majority of maize genes have been identified by at least one currently available GSS. Both methyl-filtration and high-Cot enrichment methods provided a 7- to 8-fold increase in gene discovery rates as compared to random sequencing. The available maize GSSs aligned to 75% of the FLC nucleotides used to perform searches, while the expressed sequence tag sequences aligned to 73% of the nucleotides. Our data suggest that at least approximately 95% of maize genes have been tagged by at least one GSS. While the GSSs are very effective for gene identification, relatively few (18%) of the FLCs are completely represented by GSSs. Analysis of the overlap of coverage and bias due to position within a gene suggest that RescueMu, methyl-filtration, and high-Cot methods are at least partially nonredundant.

摘要

玉米(Zea mays)拥有一个庞大的、高度重复的基因组,因此,人们使用了多种简化代表性测序方法,试图在避开与重复DNA相关困难的同时,富集基因空间。本文记录了公开可用的玉米表达序列标签和基因组调查序列(GSSs;其中许多是通过使用简化代表性技术分离得到的)识别并覆盖78个玉米全长cDNA(FLCs)的能力。数据集中的所有78个FLCs均由至少三个GSSs鉴定出来,这表明大多数玉米基因已被至少一个现有GSSs鉴定。与随机测序相比,甲基过滤和高Cot值富集方法的基因发现率提高了7至8倍。可用的玉米GSSs与用于搜索的FLC核苷酸的75%对齐,而表达序列标签序列与73%的核苷酸对齐。我们的数据表明,至少约95%的玉米基因已被至少一个GSSs标记。虽然GSSs在基因鉴定方面非常有效,但相对较少(18%)的FLCs由GSSs完全代表。对基因内覆盖重叠和位置偏差的分析表明,RescueMu、甲基过滤和高Cot值方法至少部分是非冗余的。

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