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目的 HRalGDS基因是我们新近分离与克隆的人类新的Ras相关基因,是鼠Ral鸟嘌呤核苷酸解离刺激因子(RalGDS)在人类的同源基因。利用辐射杂种板(radiationhybrid,RH)制图法进行该基因的精细定位。方法 根据HRalGDS基因cDNA的3′不翻译区设计正反向引物,PCR扩增人鼠辐射体细胞杂种板(radiationhybridGenebridge4panel),并将杂种板每一细胞株的PCR结果按一定方式统计后输入WIMITRadiationHybridMapper的相关网址,进行HRalGDS基因的RH制图和整合分析。结果RH制图将HRalGDS基因定位在9号染色体的骨架图上。定位附近的Marker顺序为9pter——WI6083——HRalGDSWI1405——9qter,经文献检索及进一步与物理图、遗传连锁图及细胞学图谱的整合分析,将其精细定位于染色体9q34.1区带的基因位标SURF5与RPL7A之间。结论 HRalGDS基因的定位不仅有助于人染色体9q34区带的精细基因图与细胞遗传图的构建,而且也表明用RH制图法定位于人类新基因既简便、易行、可靠,又可为丰富染色体上的基因定位提供新的信息。
Purpose The HRalGDS gene is our newly isolated and cloned human Ras-related gene and is a human homologue of RalGDS. Fine mapping of this gene was performed using radiationhybrid (RH) mapping. Methods Based on the 3 ’untranslated region of HRalGDS gene, forward and reverse primers were designed and PCR products were amplified by PCR. The PCR results of each cell line were statistically analyzed and entered into the WIMIT Radiation Hybrid Mapper Related URLs for HR mapping of HRGGDS genes and integration analysis. Results RH mapping mapped the HRalGDS gene on the chromosome 9 cytoskeleton map. The sequence of Marker near the target was 9pter - WI6083 - HRalGDSWI1405--9qter. After the literature search and further integration analysis with physical map, genetic linkage map and cytology map, the gene whose locus was finely localized on chromosome 9q34.1 Between SURF5 and RPL7A. Conclusion The localization of HRalGDS gene not only contributes to the construction of fine gene map and cytogenetic map of human chromosome 9q34, but also shows that the new human gene regulated by RH mapping is simple, easy and reliable, and rich in chromosomes The gene mapping provides new information.