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大熊猫(Ailuropoda melanoleuca)7倍体细菌人工染色体文库的构建及10个基因的细胞遗传学定位

Construction of a 7-fold BAC library and cytogenetic mapping of 10 genes in the giant panda (Ailuropoda melanoleuca).

作者信息

Liu Wei, Zhao Yonghui, Liu Zhaoliang, Zhang Ying, Lian Zhengxing, Li Ning

机构信息

State Key Laboratory for Agro-Biotechnology, China Agricultural University, Beijing 100094, China.

出版信息

BMC Genomics. 2006 Nov 17;7:294. doi: 10.1186/1471-2164-7-294.

Abstract

BACKGROUND

The giant panda, one of the most primitive carnivores, is an endangered animal. Although it has been the subject of many interesting studies during recent years, little is known about its genome. In order to promote research on this genome, a bacterial artificial chromosome (BAC) library of the giant panda was constructed in this study.

RESULTS

This BAC library contains 198,844 clones with an average insert size of 108 kb, which represents approximately seven equivalents of the giant panda haploid genome. Screening the library with 15 genes and 8 microsatellite markers demonstrates that it is representative and has good genome coverage. Furthermore, ten BAC clones harbouring AGXT, GHR, FSHR, IRBP, SOX14, TTR, BDNF, NT-4, LH and ZFX1 were mapped to 8 pairs of giant panda chromosomes by fluorescence in situ hybridization (FISH).

CONCLUSION

This is the first large-insert genomic DNA library for the giant panda, and will contribute to understanding this endangered species in the areas of genome sequencing, physical mapping, gene cloning and comparative genomic studies. We also identified the physical locations of ten genes on their relative chromosomes by FISH, providing a preliminary framework for further development of a high resolution cytogenetic map of the giant panda.

摘要

背景

大熊猫是最原始的食肉动物之一,属于濒危动物。尽管近年来它一直是许多有趣研究的对象,但对其基因组的了解却很少。为了推动对该基因组的研究,本研究构建了大熊猫细菌人工染色体(BAC)文库。

结果

该BAC文库包含198,844个克隆,平均插入片段大小为108 kb,约相当于大熊猫单倍体基因组的7倍。用15个基因和8个微卫星标记筛选该文库表明它具有代表性且基因组覆盖良好。此外,通过荧光原位杂交(FISH)将携带AGXT、GHR、FSHR、IRBP、SOX14、TTR、BDNF、NT - 4、LH和ZFX1的10个BAC克隆定位到大熊猫的8对染色体上。

结论

这是首个针对大熊猫的大插入片段基因组DNA文库,将有助于在基因组测序、物理图谱构建、基因克隆和比较基因组研究等领域对这一濒危物种的了解。我们还通过FISH确定了10个基因在其相对染色体上的物理位置,为进一步绘制高分辨率大熊猫细胞遗传图谱提供了初步框架。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a5fd/1664575/afa595dc271b/1471-2164-7-294-1.jpg

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