Method and kit for gene sequencing and typing of human major histocompatibility complex class I chain related gene A (MICA)

Abstract

The invention relates to a method and a kit for gene sequencing and typing of a human major histocompatibility complex class I chain related gene A (MICA). The method comprises the following specific steps: extracting sample genome DNA (deoxyribonucleic acid); designing a PCR (polymerase chain reaction) primer and carrying out PCR amplification on the sample genome DNA; designing a sequencing primer, and carrying out dual-direction sequencing on the obtained PCR amplification product, namely respectively carrying out forward and reverse dual-direction sequencing on 1,2,3,4,5 and 6 exons in the PCR amplification product; and typing according to the sequencing result. By using the method, the problem that the existing imported MICA commercialized sequencing kit is just a single-direction sequencing kit is solved, the defects that the base signal intensity of single-direction sequencing is not in consistence and the existing of mutation can be not clearly reflected are avoided, ambiguous gene typing result of MIAC is solved, and the base sequences of the first and sixth exons are obtained.

Description

【Technical field】 [0001] The invention relates to a sequencing typing method, in particular to a sequencing typing method for human major histocompatibility complex class I chain related gene A (MICA). 【Background technique】 [0002] The human major histocompatibility complex class I molecule-associated gene A gene (MICA gene) is located in the short arm of the human chromosome 6 HLA-III gene region, and belongs to the functional gene of the non-classical HLA-class I gene family. [0003] The full length of the MICA gene is 11722bp (NM_000247), encoding a transcript of 1382bp. The whole gene includes 6 exons (exons 1-6), exon 1 encodes the L leader peptide, exons 2-4 encode the extracellular α1-3 domain respectively, and exon 5 encodes the transmembrane ( TM) region, exon 6 encodes the cytoplasmic region. [0004] Table 1 Length of each exon and intron of MICA gene [0005] [0006] [0007] The current MICA genotyping methods include polymerase chain reaction seque...

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