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盘基网柄菌发育cDNA项目:发育第一指阶段表达序列标签的产生与分析

The Dictyostelium developmental cDNA project: generation and analysis of expressed sequence tags from the first-finger stage of development.

作者信息

Morio T, Urushihara H, Saito T, Ugawa Y, Mizuno H, Yoshida M, Yoshino R, Mitra B N, Pi M, Sato T, Takemoto K, Yasukawa H, Williams J, Maeda M, Takeuchi I, Ochiai H, Tanaka Y

机构信息

Institute of Biological Sciences, Univ., Tsukuba, Ibaraki, Japan.

出版信息

DNA Res. 1998 Dec 31;5(6):335-40. doi: 10.1093/dnares/5.6.335.

Abstract

In an effort to identify and characterize genes expressed during multicellular development ill Dictyostelium, we have undertaken a cDNA sequencing project. Using size-fractionated subsets of cDNA from the first finger stage, two sets of gridded libraries were constructed for cDNA sequencing. One, library S, consisting of 9984 clones, carries relatively short inserts, and the other, library L, which consists of 8448 clones, has longer inserts. We sequenced all the selected clones in library S from their 3'-ends, and this generated 3093 non-redundant, expressed sequence tags (ESTs). Among them, 246 ESTs hit known Dictyostelium genes and 910 showed significant similarity to genes of Dictyostelium and other organisms. For library L, 1132 clones were randomly sequenced and 471 non-redundant ESTs were obtained. In combination, the ESTs from the two libraries represent approximately 40% of genes expressed in late development, assuming that the non-redundant ESTs correspond to independent genes. They will provide a useful resource for investigating the genetic networks that regulate multicellular development of this organism.

摘要

为了鉴定和描述盘基网柄菌多细胞发育过程中表达的基因,我们开展了一个cDNA测序项目。利用来自第一指状期的cDNA大小分级亚群,构建了两组用于cDNA测序的网格化文库。一个是文库S,由9984个克隆组成,插入片段相对较短;另一个是文库L,由8448个克隆组成,插入片段较长。我们从文库S中所有选定克隆的3'端进行测序,共产生了3093个非冗余的表达序列标签(EST)。其中,246个EST与已知的盘基网柄菌基因匹配,910个与盘基网柄菌及其他生物的基因具有显著相似性。对于文库L,随机测序了1132个克隆,获得了471个非冗余EST。假设非冗余EST对应于独立基因,那么这两个文库的EST组合起来大约代表了发育后期表达基因的40%。它们将为研究调控该生物体多细胞发育的遗传网络提供有用的资源。

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