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Published on: February 28, 2015
[Establishment of a large-scale bi-directional sequencing and genotyping platform for MICA gene exons 2 to 4]
Su-qing Gao1, Zhi-hui Deng, Yun-ping Xu
1Shenzhen Blood Center, Shenzhen Key Laboratory of Histocompatibility and Immunogenetics, Shenzhen, Guangdong, People's Republic of China. gsq_dengzhihui@163.com
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.
Conclusion:
The bi-directional sequencing genotyping platform may be applied for large-scale study of MICA allelic polymorphisms, tissue typing, organ transplantation and disease research.
