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2025年8月18日 星期一
  2018, Vol. 26 Issue (10): 1805-1813    
  研究资源与技术改进 本期目录 | 过刊浏览 | 高级检索 |
家鸽基因分型系统的建立及应用
祝国强1,伏东科2,罗汉宋3,冯继东4,李生斌2
1. 深圳市法医学重点实验室,
2. 深圳市华大法医科技有限公司
3. 深圳市尊贵科技有限公司
4. 中国农业大学 农业生物技术国家重点实验室
Development and Application of the Genotyping System in Pigeon (Columba livia domestica)
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摘要 以短串联重复序列(short tandem repeats, STR)和性别基因为基础的动物分型系统,不仅能提供重要的分子遗传标记信息,还能有效地解决动物个体识别和亲缘关系难题。为建立家鸽(Columba livia domestica)短串联重复序列与染色体螺旋蛋白基因(chromobox-helicase-DNA binding gene, CHD)复合扩增检测体系,并评估其法医学应用价值,本研究采用六色荧光标记复合扩增方法,选取了CliμD11、PG4、PG1、PG2、CliμT02、CliμD17、CliμD35、CliμT17、CliμD16、PIGN04、CliμD32、PIGN57、PIGN26、PG5、PG6、CliμD19、PIGN12、PIGN15、PIGN10和性别鉴定基因CHD,共计20个基因位点。结果显示,本研究建立的家鸽复合扩增体系的分型结果稳定、图谱清晰,241只家鸽的羽毛样本检测全部稳定可靠;检测灵敏度高达0.0625 ng;与其他物种对比扩增,表现出高度的种属特异性;对19个STR位点的等位基因在232只家鸽群体中的统计分析表明,本研究所选取的STR位点均可用于家鸽的个体识别和亲子鉴定。本研究最终成功建立的稳定性好、灵敏度高、种属特异性强的家鸽STR位点复合扩增检测体系,是目前已知位点数最多的家鸽检测体系,也是国内相关研究的首次报道,为家鸽的个体识别和亲子鉴定提供了分子鉴定基础。
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祝国强
伏东科
罗汉宋
冯继东
李生斌
关键词 家鸽短串联重复序列微卫星复合扩增    
Abstract:The genotyping system based on short tandem repeats (STR) and sex locus can not only provide important molecular genetic marker information, but also effectively solve the problem of individual identification and genetic relationship in animals. To establish a multiplex detection system for STR and CHD (chromobox-helicase-DNA binding gene) of pigeon (Columba livia domestica) and evaluate its forensic application value, in present study, a 6-dye fluorescence-labelling multiplex PCR amplification system for 19 STR (short tandem repeats) loci and 1 CHD sex locus was established to evaluate its forensic application and population research. In total, CliμD11, PG4, PG1, PG2, CliμT02, CliμD17, CliμD35, CliμT17, CliμD16, PIGN04, CliμD32, PIGN57, PIGN26, PG5, PG6, CliμD19, PIGN12, PIGN15, PIGN10, and gender-identification gene CHD were picked up. The results showed that the multiplex amplification system was reliable and easy for identification via feather examples from 241 pigeons. It had been proved that the sensitivity could be low to 0.0625 ng DNA and the species identification was rather specific. The statistic analysis of all alleles of 19 STR loci in the pigeon population showed that all 19 STR loci could be qualified to be used in individual identification and paternity test. Finally it was proved that the pigeon STR loci multiplex amplification system established in present research was rather stable, sensitive and species-specific, which was the most effective pigeon detection system and the first report in China as well. The present study research provides molecular basis for individual identification and paternity testing in pigeons.
Key wordsPigeon    Short tandem repeat (STR)    Microsatellite    Multiplex PCR
收稿日期: 2018-06-01      出版日期: 2018-09-26
基金资助:深圳市科技计划项目资助
通讯作者: 李生斌     E-mail: silkroad@genomics.cn
引用本文:   
祝国强 伏东科 罗汉宋 冯继东 李生斌. 家鸽基因分型系统的建立及应用[J]. , 2018, 26(10): 1805-1813.
链接本文:  
http://journal05.magtech.org.cn/Jwk_ny/CN/     或     http://journal05.magtech.org.cn/Jwk_ny/CN/Y2018/V26/I10/1805
 
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