title: [Establishment of a large-scale bi-directional sequencing and genotyping platform for MICA gene exons 2 to 4].
abstract: OBJECTIVE
  To 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.
  
  
  METHODS
  Primers 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.
  
  
  RESULTS
  Results 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.
  
  
  CONCLUSIONS
  The bi-directional sequencing genotyping platform may be applied for large-scale study of MICA allelic polymorphisms, tissue typing, organ transplantation and disease research.
authors.affiliation: Shenzhen Blood Center, Shenzhen Key Laboratory of Histocompatibility and Immunogenetics, Shenzhen, Guangdong, People's Republic of China. gsq_dengzhihui@163.com
authors.forename: Su-qing
authors.initials: SQ
authors.lastname: Gao
authors.forename: Zhi-hui
authors.initials: ZH
authors.lastname: Deng
authors.forename: Yun-ping
authors.initials: YP
authors.lastname: Xu
authors.forename: Da-ming
authors.initials: DM
authors.lastname: Wang
authors.forename: Liu-mei
authors.initials: LM
authors.lastname: He
authors.forename: Shi-zheng
authors.initials: SZ
authors.lastname: Jin
mesh_terms: D000328:Adult; D001483:Base Sequence; D005091:Exons; D005260:Female; D060005:Genotyping Techniques; D015395:Histocompatibility Antigens Class I / Q000235:genetics*; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D020641:Polymorphism, Single Nucleotide; D017422:Sequence Analysis, DNA
chemical_list: D015395:Histocompatibility Antigens Class I; C099187:MHC class I-related chain A