人类GABARAPL2基因的染色体定位

Chromosomal mapping of GABARAPL2

为了确定人类GABAA 受体相关蛋白相似蛋白 2基因在染色体上的位置 ,根据该基因cDNA的 3′非翻译区序列设计一对RH定位引物 ,以人 /鼠体细胞辐射杂种板Genebridge4(GB4) panel和G3 panel试剂盒中的杂种细胞株基因组DNA为模板 ,在一定的条件下进行PCR扩增 ,琼脂糖凝胶电泳结果分别输入Sanger和Stanford网站的RH定位分析系统进行统计学分析。结果PCR法在一些杂种细胞株中扩增出特异的目的片段 ,并经测序验证了其准确性。凝胶电泳结果在Sanger网站统计分析表明该基因与 16号染色体上的AFM 3 4 0 ye5 ,AFM 2 92xh5 ,AFM 3 2 0wf1,AFMa0 66xd5 ,AFM 2 49xc5位标紧密连锁 ;在Stanford网站统计分析表明该基因与 16号染色体上的SHGC— 14 5 7,SHGC— 5 3 4 15 ,SHGC— 6782 ,SHGC— 2 2 2 8,SHGC— 14 62 9位标紧密连锁 ,LOD值均大于 3。整合分析该染色体区带的物理图、遗传图及细胞图谱后 ,最终将基因定位在染色体 16q2 2 .3区带的D16s3 0 16和D16s5 15位标之间。结论放射杂交定位法是一种新颖便捷的基因定位的方法 ,通过此法成功地进行了人类GABAA 受体相关蛋白相似蛋白 2基因的定位。

In order to locate GABARAPL2(GABA A-receptor-associated protein like 2)in human chromosome,a pair of primers were designed according to the 3′ untranslated region sequence of its cDNA.PCR were performed under certain conditions by using hybrids genomic DNA in Genebridge 4(GB4) panel and G3 panel as templates.The electrophoresis results on agarose gel were sent to Sanger RH Mapping Server and Stanford RH Mapping Server for statistical analysis.Results The predicted fragments amplified by PCR were confirmed by sequencing.It was showed that the gene is tightly linked with markers AFM340ye5,AFM292xh5,AFM320wf1,AFMa066xd5 and AFM249xc5 with GB4 panel,and that the gene is tightly linked with markers SHGC-1457,SHGC-53415,SHGC-6782,SHGC-2228,SHGC-14629 on chromosome 16 with G3 panel.Lod score>3 under two circumstances.The integrated analyses of comprehensive map,cytogenetic map in Genomic Database of the gene indicated that GABARAPL2 was assigned to chromosome 16q22.3 between markers D16s3016 and D16s515.Conclusion Radiation Hybrid Mapping method is a new and convenient method to map genes.With this method,GABARAPL2 is successfully mapped to chromosome 16q22.3 in this paper.