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首页> 外文期刊>Mammalian genome: official journal of the International Mammalian Genome Society >RH mapping of canine TOAST markers: a new strategy for species-specific primer design to prevent amplification of host orthologous gene products especially with similar sizes
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RH mapping of canine TOAST markers: a new strategy for species-specific primer design to prevent amplification of host orthologous gene products especially with similar sizes

机译：犬TOAST标记的RH定位：物种特异性引物设计的一种新策略，可防止宿主直系同源基因产物的扩增，尤其是大小相似的产物

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The high degree of sequence conservation between orthologous genes has allowed development of reagents for mammalian comparative gene mapping and amplification of orthologous genes in different species. Traced orthologous amplified sequence tags (TOASTs; Jiang et al. 1998), comparative anchor-tagged sequences (CATS; Lyons et al. 1997), and gene-specific universal mammalian sequence-tagged sites (Venta et al. 1996) have been used successfully to transfer gene sequence information from maprich species to map-poor species. The specific feature of these markers is that their primers are derived within conserved regions of orthologous gene sequences among different mammals. As radiation hybrid mapping is a method of choice to map genes in a given genome, a significant problem often raised is the coamplification of the orthologous gene products with similar sizes in both donor and host species when using such "universal" primers. For example, out of the 139 pairs of TOAST primers that successfully amplified the canine gene products, 91 (66 percent) can also amplify the hamster gene products (Priat et al. 1999), and finally only 25 have been mapped on a dog radiation hybrid panel (Priat et al. 1998; Vignaux et al. 1999). We even used the canine TOAST sequences to derive primers, and 48 percent (13/27) of them can still amplify hamster gene products with similar sizes (Priat et al. 1999). Clearly, co-amplification of donor and host gene products with similar sizes is a significant limitation for using the genespecific universal primers directly in RH mapping.
机译：直系同源基因之间高度的序列保守性允许开发用于哺乳动物比较基因作图和扩增不同物种中直系同源基因的试剂。使用了追踪的直系同源扩增序列标签（TOASTs; Jiang等，1998），比较锚定标签的序列（CATS; Lyons等人，1997）和基因特异性通用哺乳动物序列标签的位点（Venta等，1996）。成功地将基因序列信息从地图丰富的物种转移到地图贫乏的物种。这些标志物的特定特征是它们的引物来源于不同哺乳动物直系同源基因序列的保守区内。由于辐射杂交作图是在给定基因组中作图的一种选择方法，因此经常出现的一个重要问题是当使用这种“通用”引物时，在供体和宿主物种中共扩增大小相似的直系同源基因产物。例如，在成功扩增犬类基因产物的139对TOAST引物中，有91对（66％）还可以扩增仓鼠基因产物（Priat等，1999），最后只有25条在狗辐射中定位混合小组（Priat等人1998; Vignaux等人1999）。我们甚至使用犬TOAST序列来获得引物，其中48％（13/27）仍可以扩增相似大小的仓鼠基因产物（Priat et al。1999）。显然，将相似大小的供体和宿主基因产物共同扩增是直接在RH作图中直接使用基因特异性通用引物的重大限制。

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