Establishment of a large-scale bi-directional sequencing and genotyping platform for MICA gene exons 2 to 4.
- VernacularTitle:MICA基因第2~4外显子大规模双向测序及分子克隆检测平台的建立
- Author:
Su-qing GAO
1
;
Zhi-hui DENG
;
Yun-ping XU
;
Da-ming WANG
;
Liu-mei HE
;
Shi-zheng JIN
Author Information
- Publication Type:Journal Article
- MeSH: Adult; Base Sequence; Exons; Female; Genotyping Techniques; Histocompatibility Antigens Class I; genetics; Humans; Male; Molecular Sequence Data; Polymerase Chain Reaction; Polymorphism, Single Nucleotide; Sequence Analysis, DNA
- From: Chinese Journal of Medical Genetics 2012;29(5):542-546
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo establish a stable and large-scale bi-directional sequencing platform for genotyping MICA gene exons 2 to 4, and to analyze single nucleotide polymorphisms(SNP) of the region.
METHODSPrimers for particular alleles of MICA gene exons 2 to 5 were designed. Optimal conditions for PCR amplification and sequencing reaction were explored. A commercialized one-way sequencing kit for MICA allele was used as a parallel control. Four samples carrying a MICA *010 allele were subjected to cloning and haplotype sequencing.
RESULTSResults of MICA allele typing of 100 samples for a parallel control group were confirmed by the establish method. Twenty-two SNP in MICA gene exons 2 to 4 were detected in Chinese population. Two novel allelic sequences were accepted by GenBank and IMGT/HLA database and officially named as MICA*065 and MICA*066 by the WHO Nomenclature Committee. A novel SNP in MICA gene intron 3 was discovered, with allelic sequence submitted to GenBank and IMGT/HLA database.
CONCLUSIONThe bi-directional sequencing genotyping platform may be applied for large-scale study of MICA allelic polymorphisms, tissue typing, organ transplantation and disease research.