KIR and ligand genetic typing experimental method

Abstract

The invention discloses a KIR and ligand genetic typing experimental method. By the technology, the time and labor are saved, and meanwhile, DNA sample capacity is further saved. A multi-PCR technology is adopted, meanwhile, 36 pairs of primers are further combined according to the sizes of PCR products, and finally, amplified reaction is finished in 12 reaction holes. The KIR and ligand genetic typing experimental method is conveniently applied to amplification of 96 pore plates, conventional Taq enzyme is used, typing of all KIR genes and ligands thereof can be finished at a time under the same reaction conditions, and the practicality of the KIR and ligand genetic typing experimental method is greatly improved. Two pairs of primers are used for a KIR gene, the accuracy is improved, anda false negative result is avoided. The KIR gene can be combined to MHC-I type ligand molecules on the surfaces of targeting cells, inhibiting or activating signals are transmitted to regulate the activity of NK cells and T cells, and the KIR and ligand genetic typing experimental method plays an important regulation role in hematopoietic stem cell transplantation, feto-matemal tolerance, anti-infectious immunity, tumor immunity and autoimmune diseases. Therefore, KIR genetic typing facilitates understanding of influences of KIR to tumor immunity, hematopoietic stem cell transplantation and autoimmune diseases.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a KIR and ligand genotyping experimental method. Background technique [0002] Natural killer cells (NK) are a kind of "bridge" cells connecting innate immunity and acquired immunity. The function of NK cells is regulated by different receptors, among which an important class of receptors is called killer immunoglobulin-like receptors (KIR). KIR is a series of molecules belonging to the immunoglobulin-like superfamily, expressed on the surface of NK cells and some T cells. [0003] With the continuous discovery of new KIR genes and alleles and the continuous application of new technologies, polymerase chain reaction-sequence-specific primers (PCR-SSP) and polymerase chain reaction-sequence-specific oligonucleotides have been established so far. acid probe (PCR-SSOP), polymerase chain reaction-direct sequencing typing (PCR-SBT) and other methods. [0004] 1. PCR-SSP [0005] Uhrbe...

AI Technical Summary

Problems solved by technology

However, since most of the reaction primers select to amplify exons encoding immunoglobulin-like domains, and these exons are separated by about 1.5kbp introns, the amplified products of the above PCR reactions are mostly long fragments with a size 0.5-2.0kbp, amplification requires high-quality DNA samples

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