Specific primer and liquid chip for detecting polymorphism of NAT1 (N-acetyltransferase 1) gene

Abstract

The invention discloses a liquid chip and specific primer for detecting an NAT1 (N-acetyltransferase 1) gene. The liquid chip mainly comprises ASPE (allele specific primer extension) primers, microspheres and amplification primers, wherein each ASPE primer is formed by tag sequences at the 5' terminal and specific primer sequences which are arranged at the 3' terminal and aim at the mutation site of the target gene; the specific primer sequences are SEQ ID NO.15 and SEQ ID NO.16 aiming at the C135T site, SEQ ID NO.17 and SEQ ID NO.18 aiming at the G136A site, SEQ ID NO.19 and SEQ ID NO.20 aiming at the T216G site, SEQ ID NO.21 and SEQ ID NO.22 aiming at the A328T site, SEQ ID NO.23 and SEQ ID NO.24 aiming at the A460T site, SEQ ID NO.25 and SEQ ID NO.26 aiming at the C106T site and / or SEQ ID NO.27 and SEQ ID NO.28 aiming at the C199T site; and the microspheres are enveloped by anti-tag sequences. The detecting liquid chip provided by the invention has the advantages that the consistency of the detection result and the sequencing method is as high as 100%, thus parallel detection of wild types and mutant types of a plurality of mutation sites is achieved.

Description

technical field [0001] The invention belongs to the field of molecular biology, relates to medicine and biotechnology, in particular to a specific primer for NAT1 gene polymorphism detection and a liquid phase chip. Background technique [0002] N-acetylation metabolism is an important biotransformation pathway for many arylamines and hydrazines, catalyzed by N-acetyltransferase (NAT) dependent on acetyl-CoA, N-acetyltransferase (N-acetyltrans ferasel , NAT) is an enzyme system that catalyzes the acetylation of nitrogen-containing compounds in the body. It can activate or degrade nitrogen-containing exogenous substances, especially exogenous carcinogens containing amine groups. The genes encoding NAT are NAT1 and NAT2, which are located on the 8th pair of chromosomes (8p21.3-23.1). Studies have shown that human NAT1 has a wider tissue distribution and exists in colonic mucosa, liver, bladder, red blood cells and lymphocytes , is the dominant acetyltransferase in colorectal ...

AI Technical Summary

Problems solved by technology

The PCR-RFLP method is based on the change of the restriction endonuclease recognition site caused by gene mutation, such as the loss of the site or the generation of a new site, a specific fragment is amplified by PCR, and then cut and amplified with a restriction endonuclease. Amplify the product and observe the size of the fragment by electrophoresis. This method is used to detect gene mutations with altered restriction sites, and can directly determine the genotype. However, this method cannot be used for the detection of gene mutations without new restriction sites.

Again, the above methods all have limitations in detection throughput, and can only detect one mutation type at a time, which cannot meet the needs of practical applications.

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