人类GABARAPL1基因的染色体定位

Chromosomal Mapping of Human GABARAPL1

目的 确定人类GABAA 受体相关蛋白相似蛋白 1基因在染色体上的位置。方法 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Genebridge 4(GB4) panel试剂盒中的杂种细胞株基因组DNA为模板 ,进行PCR扩增 ,经琼脂糖凝胶电泳检测后 ,按Sanger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 ,经测序验证了该序列的准确性。通过Sanger网站统计分析表明 ,该基因与 12号染色体上的AFM 0 2 6tb5 (D12S77) ,AFM32 0xb5 (D12S35 8) ,AFM 2 0 5xg3(D12S310 ) ,AFM2 17xa7(D12S99) ,AFM 2 34wb6 (D12S16 6 4)位标紧密连锁 ,且LOD值均大于 3。经该染色体区带的物理图、遗传图及细胞图谱整合分析 ,将该基因定位在染色体 12 p12 .3区带的D12S77和D12S35 8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 1基因的染色体定位。

Objective To map the GABARAPL1 (GABA A-receptor-associated protein like 1) gene in human chromosome.Methods A pair of primers were designed according to the 3′ untranslated region sequence of the cDNA of GABARAPL1. PCR was performed under certain conditions by using hybrids genomic DNA in Genebridge 4 (GB4) panel as templates. The electrophoresis results on agarose gel were sent to Sanger RH Mapping Server for statistical analysis.Results The predicted fragments were amplified by PCR and were then confirmed by sequencing. The analysis in Sanger RH Mapping Server showed that the gene was tightly linked with markers AFM026tb5 (D12S77),AFM320xb5 (D12S358),AFM205xg3 (D12S310),AFM217xa7 (D12S99),AFM234wb6 (D12S1664) on chromosome 12. Lod score was higher than 3. The integrated analyses of comprehensive map, cytogenetic map in Genomic Database of the gene indicated that GABARAPL1 was assigned to chromosome12p12.3 between markers D12S77 and D12S358.Conclusion Radiation Hybrid Mapping method is a new and convenient method to map genes. With this method, human GABARAPL1 is successfully mapped to chromosome12p12.3.