A somatic cell hybrid panel for pig regional gene mapping characterized by molecular cytogenetics
A somatic cell hybrid panel for pig regional gene mapping characterized by molecular cytogenetics
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DOI：
10.1159/000134338
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发表时间：
1996-01-01
期刊：
影响因子：
--
通讯作者：
Gellin, J
中科院分区：
文献类型：
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作者：
Yerle, M;Echard, G;Gellin, J
产生了一组27头猪x啮齿动物体细胞杂种，并进行了细胞遗传学表征。本研究的第一步是将SINE探针与每个杂交克隆的GTG带中期分裂相杂交，以计数和鉴定正常的猪染色体并检测重排的染色体。第二次渗漏包括在pIRS-PCR（猪散布重复序列-聚合酶链反应）扩增后使用每个克隆的DNA作为探针，以在猪序列中高度富集它。这些探针，杂交到正常的猪中期染色体，使杂交系中的完整的猪补体的鉴定。整个染色体和片段的特点是快速，准确，结果进行了比较。除了这种细胞遗传学特征，还进行了分子验证，通过使用特异性引物的6个微卫星和一个基因先前映射到猪染色体。获得的结果使我们能够得出结论，我们已经产生了一个面板，是所有猪染色体的信息。该小组构成了一个高效的工具，不仅可以确定基因和标记的分配，还可以确定猪染色体上的区域定位。
A panel of 27 pig x rodent somatic cell hybrids was produced and characterized cytogenetically. The first step of this study consisted of hybridizing a SINE probe to GTG-banded metaphases of each hybrid clone in order to count and identify the normal pig chromosomes and to detect rearranged ones. The second seep consisted of using the DNA of each clone as a probe after pIRS-PCR (porcine interspersed repetitive sequence-polymerase chain reaction) amplification to highly enrich it in pig sequences. These probes, hybridized to normal pig metaphase chromosomes, enabled the identification of the complete porcine complement in the hybrid lines. Whole chromosomes and fragments were characterized quickly and precisely, and results were compared. In addition to this cytogenetic characterization, molecular verification was also carried out by using primers specific to six microsatellites and to one gene previously mapped to pig chromosomes. The results obtained allow us to conclude that we have produced a panel that is informative for all porcine chromosomes. This panel constitutes a highly efficient tool to establish not only assignments of genes and markers but also regional localizations on pig chromosomes.