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  

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

作　　者：郑其平[1] 余龙[1] 赵勇[1] 孙喜元 戴方彦[1] 胡培蓉[1] 付强[1] 庚镇城[1] 

机构地区：[1]复旦大学遗传研究所遗传工程国家重点实验室,上海200433

出　　处：《中华医学遗传学杂志》2000年第1期1-5,共5页Chinese Journal of Medical Genetics

基　　金：863计划资助项目!Z19020401;国家杰出青年基金!3952 0 51 5;上海市科技发展重大项目!96JC1 4 0 0 1

摘　　要：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。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 infomation of genes within chromsomal regions.

关 键 词：H-RalGDS 基因 辐射杂种板制图 染色体9q34.1 

分 类 号：R394[医药卫生—医学遗传学]