| Transcribed single nucleotide polymorphism: Ideal markers for detecting gene imprinting by 5' nuclease assay |
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| 作者姓名： | 朱冠山 万谟彬 朱忠政 郑瑞英 |
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| 作者单位： | Department of Infectious Diseases，Changhai Hospital，Second Military Medical University，Shanghai 200433，China National Institutes of Health，Maryland 20850，USA，Department of Infectious Diseases，Changhai Hospital，Second Military Medical University，Shanghai 200433，China，Eastern Hepatobiliary Hospital，Second Military Medical University，Shanghai 200433，China，Department of Infectious Diseases，Changhai Hospital，Second Military Medical University，Shanghai 200433，China |
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| 摘 要： | Objective: To establish a novel approach for quick and highly efficient verification of human gene imprinting. Methods: A pair of dye-labelled probes, 5' nuclease assay was combined with RT-PCR to determine the genotype of a transcribed single nucleotide polymorphism (SNP) rs705(C>T) of a known imprinted gene, small nuclear ribonucleotide protein N (SNRPN), on both genomic DNA and cDNA of human lym-phoblast cell lines. Results: Allele discrimination showed a clear monoallelic expression pattern of SNRPN, which was confirmed by RT-PCR based restriction fragment length polymorphism (RFLPs). Pedigree analysis verified the paternal origin of expressed allele, which was in consistency with previous report. Conclusion: Transcribed SNP is an ideal marker for detecting gene imprinting by 5' nuclease assay. This approach also may be used to discover differential allele expression of non-imprinted genes, finding out gene cis-acting functional polymorphism.
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Transcribed single nucleotide polymorphism: Ideal markers for detecting gene imprinting by 5' nuclease assay |
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| ZHU Guan-Shan,WAN Mo-Bin,ZHU Zhong-zheng,ZHENG Rui-ying.Transcribed single nucleotide polymorphism: Ideal markers for detecting gene imprinting by 5' nuclease assay[J].Journal of Medical Colleges of PLA(China),2002,17(4). |
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| Authors: | ZHU Guan-Shan WAN Mo-Bin ZHU Zhong-zheng ZHENG Rui-ying |
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| Abstract: | Objective: To establish a novel approach for quick and highly efficient verification of human gene imprinting. Methods: A pair of dye-labelled probes, 5' nuclease assay was combined with RT-PCR to determine the genotype of a transcribed single nucleotide polymorphism (SNP) rs705(C>T) of a known imprinted gene, small nuclear ribonucleotide protein N (SNRPN), on both genomic DNA and cDNA of human lym-phoblast cell lines. Results: Allele discrimination showed a clear monoallelic expression pattern of SNRPN, which was confirmed by RT-PCR based restriction fragment length polymorphism (RFLPs). Pedigree analysis verified the paternal origin of expressed allele, which was in consistency with previous report. Conclusion: Transcribed SNP is an ideal marker for detecting gene imprinting by 5' nuclease assay. This approach also may be used to discover differential allele expression of non-imprinted genes, finding out gene cis-acting functional polymorphism. |
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| Keywords: | single nucleotide polymorphism genomic imprinting 5' nuclease assay |
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