[Coding single nucleotide polymorphism is an ideal marker for detecting gene imprinting by 5' nuclease assay].
OBJECTIVE
To establish a novel approach for quick and high throughput verification of human gene imprinting.


METHODS
By use of a pair of dye-labeled probes, 5' nuclease assay was combined with reverse transcriptase-PCR(RT-PCR) to genotype a coding single nucleotide polymorphism (cSNP), rs705(C/T) of a known imprinted gene, small nuclear ribonucleotide protein N (SNRPN), on both genomic DNA and cDNA of human lymphoblast cell lines.


RESULTS
Allele discrimination showed a clear monoallelic expression pattern of SNRPN, which was confirmed by RT-PCR based restriction fragment length polymorphisms. Pedigree analysis verified the paternal origin of expressed allele, which is in consistency with previous report.


CONCLUSIONS
Coding SNP is an ideal marker for detecting gene imprinting by 5' nuclease assay. This approach has also a potentiality to discover differential allele expression of non-imprinted genes in order to find gene cis-acting functional polymorphism.
Wan
Mo-bin
MB
Department of Infectious Diseases, Changhai Hospital, Second Military Medical University, Shanghai, 200433 PR China. wanmb@online.sh.cn
Zhu
Guan-shan
GS
Zheng
Rui-ying
RY
D000483:Alleles; D015415:Biomarkers; D019411:Clinical Laboratory Techniques; D004247:DNA; D004720:Endonucleases / Q000378:metabolism*; D005821:Genetic Techniques; D018392:Genomic Imprinting / Q000235:genetics*; D006801:Humans; D010375:Pedigree; D011110:Polymorphism, Genetic; D020641:Polymorphism, Single Nucleotide; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*
D015415:Biomarkers; D004247:DNA; D004720:Endonucleases