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引物原位标记技术检测SRY基因
引用本文:聂涌,丁显平,邓莉,魏萍,王欢.引物原位标记技术检测SRY基因[J].中华医学遗传学杂志,2008,25(6).
作者姓名:聂涌  丁显平  邓莉  魏萍  王欢
作者单位:四川大学生命科学学院遗传医学研究所/生物资源与生态环境教育部重点实验室,成都,610064
摘    要:目的 建立一种能更有效检测单拷贝基因的引物原位标记技术(primed in situ labeling,PRINS).方法 在传统PRINS技术基础上,引入TsqStatrt抗体、Y染色体上的性别决定区基因(SIBX determining region Y,SRY)4条引物和TSATM Biotin System来检测SRY基因,并以对SRY基因的荧光原位杂交(fluorescence in situ hybridization,FISH)为对照.结果 在PRINS结果中,通过对50个中期分裂相的分析,在Yp11.3的位置上都枪测到特异信号且清晰可辨别,此结果与FISH结果一致且标记信号强度相当.结论 这项改进后的PRINS技术可快速针对单拷贝基因和小DNA片段进行检测.因此,相对于昂贵且费时的FISH技术,是另一种有效的选择.

关 键 词:引物原位标记  SRY基因  单拷贝基因  荧光原位杂交

Detection of the SRY gene by primed in situ labeling
NIE Yong,DING Xian-ping,DENG Li,WEI Ping,WANG Huan.Detection of the SRY gene by primed in situ labeling[J].Chinese Journal of Medical Genetics,2008,25(6).
Authors:NIE Yong  DING Xian-ping  DENG Li  WEI Ping  WANG Huan
Abstract:Objective Toestablish a primed in situ labeling(PR1NS) technique which can be more effective in detection of single copy gent.Methods On the basis of traditional PRINS,new reagents and procedures,such as TaqStart antibody,four primers of the sex determining region Y (SRY) gene and TSA TM Biotin System were included in detection of the SRY gene.Meanwhile,fluorescence in situ hybridization(FISH)to detect the SRY gene was used as control.Results Fifty metaphases were scored.PRINS labeling showed signals for the SRY on the Y chromosome at band Yp11.3 in all metaphases.These signals were as distinct as that from resuhs of FISH.Conclusion This improved method is ideal for rapidly localizing single copy genes and small DNA segments.And PRINS is a cost-and time-effective alternative to FISH.
Keywords:primed in situ labeling  SRY gene  single copy gene  fluorescence in situ hybridization
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