利用辐射杂种板(GB4)精细定位人类新基因H-RalGDS于染色体9q34.1

目的 Ｈ－ＲａｌＧＤＳ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（ｒａｄｉａｔｉｏｎ ｈｙｂｒｉｄ，ＲＨ）制图法进行该基因的精细定位。方法 根据Ｈ－ＲａｌＧＤＳ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（ｒａｄｉａｔｉｏｎ ｈｙｂｒｉｄ Ｇｅｎｂｒｉｄｇｅ ４ ｐａｎｅｌ），并

Refined chromosome assignment of human novel H-RalGDS gene on chromosome 9q34.1 by using radiation hybrid Genebridge 4 panel

OBJECTIVE: Refined chromosome assignment of a novel Ras-related H-RalGDS gene (the human homology of rat RalGDS, Ral guanine nucleotide dissociation stimulator recently cloned and characterized by our laboratory) by using human/rodent radiation somatic cell hybrid panel GB4. METHODS: According to the sequence of the 3'-untranslated region of H-RalGDS cDNA, forward and reverse primers were designed and used to amplify the human/rodent somatic radiation hybrid Genebridge 4 panel. The PCR result of each cell line of the panel was scored in a certain way and put into the related internet site of WI/MIT Radiation Hybrid Mapper for RH mapping and comprehensive analysis. RESULTS: The H-RalGDS gene was successfully localized to the framework of chromosome 9. After referring to literature and further integrated mapping analysis of the physical, genetic linkage and cytological mappings, the gene was assigned precisely between the markers SURF5 and RPL7A which had already been assigned to human chromosome 9q34.1. CONCLUSION: The localization of H-RalGDS not only contributed much to the construction of refined gene map and cytogenetic map within the region of chromosome 9q34, but also made clear that RH mapping of human novel genes is a simple, convenient and reliable method which can provide even more genetic information of genes within chromosomal regions.