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用于猪精细定位的12000拉德辐射杂种细胞系的构建与鉴定

Generation and characterization of a 12,000-rad radiation hybrid panel for fine mapping in pig.

作者信息

Yerle M, Pinton P, Delcros C, Arnal N, Milan D, Robic A

机构信息

Institut National de la Recherche Agronomique, Laboratoire de Génétique Cellulaire, Castanet-Tolosan, France.

出版信息

Cytogenet Genome Res. 2002;97(3-4):219-28. doi: 10.1159/000066616.

Abstract

We have constructed a 12,000-rad porcine whole-genome radiation hybrid panel to complement the first generation 7,000-rad panel (IMpRH) and allow higher resolution mapping studies both in specific areas of interest and on the whole genome. We analyzed 243 hybrid clones on the basis of their marker retention frequency to produce a final panel of 90 hybrid clones with an average retention frequency of 35.4%. The resolution of this 12,000-rad panel (IMNpRH2) was compared to the resolution of the 7,000-rad panel (IMpRH) by constructing framework maps in the 2.4-Mb region of porcine chromosome 15 containing the acid meat RN gene. In this region, two-point analysis was used to estimate RH distances and demonstrates their reliability with the estimation of physical distances. This study demonstrates that the 12,000-rad panel constitutes a powerful tool for constructing high-resolution maps. Indeed, the resolution of IMNpRH2 (12-14 kb/cR(12,000)) is two to three times more than that of IMpRH (35-37 kb/cR(7,000)). As expected, the increase in the radiation dose allows an increase of the mapping resolution in terms of kb/cR with the same suppleness of use for mapping experiments. In addition the RH map constructed in the region investigated proved to be more homogeneous on IMNpRH2 than on IMpRH.

摘要

我们构建了一个12,000拉德的猪全基因组辐射杂种细胞系,以补充第一代7,000拉德的细胞系(IMpRH),并允许在特定感兴趣区域和全基因组上进行更高分辨率的图谱研究。我们根据标记保留频率分析了243个杂种克隆,最终得到了一个由90个杂种克隆组成的细胞系,其平均保留频率为35.4%。通过在猪15号染色体包含酸肉RN基因的2.4兆碱基区域构建框架图谱,将这个12,000拉德细胞系(IMNpRH2)的分辨率与7,000拉德细胞系(IMpRH)的分辨率进行了比较。在该区域,采用两点分析来估计辐射杂种距离,并通过物理距离估计证明了其可靠性。这项研究表明,12,000拉德细胞系是构建高分辨率图谱的有力工具。实际上,IMNpRH2(12,000拉德时为12 - 14千碱基/厘雷)的分辨率是IMpRH(7,000拉德时为35 - 37千碱基/厘雷)的两到三倍。正如预期的那样,辐射剂量的增加使得在kb/cR方面图谱分辨率提高,同时在图谱实验中具有相同的使用灵活性。此外,在所研究区域构建的辐射杂种图谱在IMNpRH2上比在IMpRH上更加均匀。

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