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构建和鉴定鲤鱼(Cyprinus carpio L.)BAC 文库及其与斑马鱼(Danio rerio)的微同线性关系的建立。

Construction and characterization of the BAC library for common carp Cyprinus carpio L. and establishment of microsynteny with zebrafish Danio rerio.

机构信息

The Centre for Applied Aquatic Genomics, Chinese Academy of Fishery Sciences, Beijing 100141, China.

出版信息

Mar Biotechnol (NY). 2011 Aug;13(4):706-12. doi: 10.1007/s10126-010-9332-9. Epub 2010 Nov 19.

Abstract

A bacterial artificial chromosome (BAC) library of common carp Cyprinus carpio L. was constructed as a part of ongoing common carp genome project, which is aiming assembly of common carp genome. The library, containing a total of 92,160 BAC clones with an average insert size of 141 kb, was constructed into the restriction site of Hind III on BAC vector CopyControl pCC1BAC, covering 7.7 X haploid genome equivalents. Three dimension pools and superpools of the BAC library were established and 23 positive clones of 14 targets were identified from one-fifth of the BAC library. Pilot project of BAC end sequencing was conducted on 2,688 BAC ends from 1,344 clones and harvested 2,522 high-quality Q20 sequences with average length of 677 bp. The sequencing success rate was 93.8% and pair-end success rate was 92.3%. A total of 212 microsyntenies had been established between common carp and zebrafish genomes as a trial for genome-wide comparative genomics in these two closely related species.

摘要

鲤鱼(Cyprinus carpio L.)的细菌人工染色体(BAC)文库作为正在进行的鲤鱼基因组项目的一部分而构建,该项目旨在组装鲤鱼基因组。该文库共包含 92160 个 BAC 克隆,平均插入大小为 141kb,构建到 BAC 载体 CopyControl pCC1BAC 的 Hind III 限制位点上,覆盖了 7.7X 单倍体基因组当量。建立了 BAC 文库的三维池和超级池,并从五分之一的 BAC 文库中鉴定出 14 个目标的 23 个阳性克隆。对来自 1344 个克隆的 2688 个 BAC 末端进行了 BAC 末端测序试点项目,收获了 2522 个高质量的 Q20 序列,平均长度为 677bp。测序成功率为 93.8%,双端测序成功率为 92.3%。共建立了 212 个微同线性群,作为这两个密切相关物种全基因组比较基因组学的试验。

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