Establishment of a large-scale bi-directional sequencing and genotyping platform for MICA gene exons 2 to 4 / 中华医学遗传学杂志
Chinese Journal of Medical Genetics
;
(6): 542-546, 2012.
Article
in Chinese
| WPRIM
| ID: wpr-232260
ABSTRACT
<p><b>OBJECTIVE</b>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.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSION</b>The bi-directional sequencing genotyping platform may be applied for large-scale study of MICA allelic polymorphisms, tissue typing, organ transplantation and disease research.</p>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Molecular Sequence Data
/
Base Sequence
/
Histocompatibility Antigens Class I
/
Polymerase Chain Reaction
/
Exons
/
Sequence Analysis, DNA
/
Polymorphism, Single Nucleotide
/
Genotyping Techniques
/
Genetics
Limits:
Adult
/
Female
/
Humans
/
Male
Language:
Chinese
Journal:
Chinese Journal of Medical Genetics
Year:
2012
Type:
Article
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