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| 印记基因5个SNP分型及亲代来源的检测 | |
| 引用本文： | 王伟妮,杨雅冉,严江伟,刘雅诚.印记基因5个SNP分型及亲代来源的检测[J].中国法医学杂志,2012,27(6):441-444,530. |
| 作者姓名： | 王伟妮 杨雅冉 严江伟 刘雅诚 |
| 作者单位： | 1. 山西医科大学法医学院,山西太原,030001 2. 中国科学院北京基因研究所,北京,100029 3. 北京市公安局法医检验鉴定中心,北京,100192 |
| 基金项目： | 国家自然科学基金项目(81172909,81072511);上海市法医学重点实验室开放课题资助项目(KF1001) |
| 摘 要： | 目的 采用复合PCR-Snapshot联合甲基化敏感酶切技术,检测印记基因中5个SNP的甲基化状态、印记亲代来源及分型.方法 选择15例亲子鉴定已证实为亲生关系的家系样本,采用单碱基延伸复合检测技术,检测家系样本IGF2AS rs1003483、SNURF rs220028、SNURF rs4906939、DLGAP2 rs6558478、SIM2 rs737380等5个SNP分型,同时选用核酸内切酶(McrBC)和甲基化敏感的限制酶(msRE) HhaⅠ、HpaⅡ消化子代DNA,验证印记基因的亲代来源.结果 经用本文方法检测,证实rs1003483为父源印记；rs220028、rs4906939为母源印记；rs6558478及rs737380未在差异甲基化区,不能确定其印记亲代来源.结论 复合PCR-Snapshot联合甲基化敏感酶切技术简单、高效,在检测多个SNP分型的同时可确定亲代来源,可在相关研究和实践中选用. |
| 关 键 词： | 法医物证学 印记基因 甲基化标记 单核苷酸多态性 |
Study on the application of parent-oforigin specific DNA methylation markers to forensic science |
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| Wang Weini,Yang Yaran,Yan Jiangwei,Liu Yacheng.Study on the application of parent-oforigin specific DNA methylation markers to forensic science[J].Chinese Journal of Forensic Medicine,2012,27(6):441-444,530. | |
| Authors: | Wang Weini Yang Yaran Yan Jiangwei Liu Yacheng |
| Institution: | 1.School of Forensic Medicine,Shanxi Medical University,Taiyuan 030001,China;2.Beijing Institute of Genomics,Beijing 100029,China;3.The Forensic Medical Examination Center,Beijing Breau of Public Security,Beijing 100192,China) |
| Abstract: | Objective To establish a methylation-sensitive enzyme predigestion combine PCR-Snapshot method for SNP genotyping and detecting DNA methylation status in imprinted genes simultaneously.Methods Five SNPs in four imprinted genes(IGF2AS rs1003483,SNURF rs220028,SNURF rs4906939,DLGAP2 rs6558478,SIM2 rs737380) were genotyped by minisequencing after multiplex PCR,and the genomic DNA of 15 blood samples from five trio cases with proven paternity was pretreated with both methylation-sensitive enzyme(HpaII,HhaI) and endonuclease McrBC to determine the parental origin of alleles.Results For the rs1003483,the result shows paternally imprinted and maternally expressed while rs220028,rs4906939show maternally imprinted and paternally expressed.rs6558478 and rs737380 can not be determined their parental origin for not in differentially methylated regions.Conclusion Our results demonstrated that Predigestion combine PCR-Snapshot method is simple,sensitive and effective for SNP genotyping and discriminating the parental origin of alleles simultaneously,which can be applied in forensic science. |
| Keywords: | forensic biological evidence imprinted genes methylation markers SNPs |
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