CYP1A1 gene polymorphism detection specific primers and liquid chip

Abstract

The invention discloses CYP1A1 gene detection specific primers and a liquid chip. The liquid chip mainly comprises: various ASPE primers composed of 5' terminal tag sequence and 3' terminal specific primer sequences aiming at target gene mutation site, microspheres coated with anti-tag sequence, and amplification primers. The specific primer sequences comprise: SEQ ID NO.17 and SEQ ID NO.18 aiming at T109C site; SEQ ID NO.19 and SEQ ID NO.20 aiming at T97C site; SEQ ID NO.21 and SEQ ID NO.22 aiming at 133-134insT site; SEQ ID NO.23 and SEQ ID NO.24 aiming at T201A site; SEQ ID NO.25 and SEQ ID NO.26 aiming at C240A site; SEQ ID NO.27 and SEQ ID NO.28 aiming at A242G site; SEQ ID NO.29 and SEQ ID NO.30 aiming at C248A site; and / or SEQ ID NO.31 and SEQ ID NO.32 aiming at G135T site. The matching rate of the detection result obtained by using the liquid chip provided by the invention and a result obtained by using a sequencing method is 100%. Therefore, wild-type and mutant-type parallel detections of a plurality of mutation sites are realized.

Description

technical field [0001] The invention belongs to the field of molecular biology, relates to medicine and biotechnology, in particular to a CYP1A1 gene polymorphism detection specific primer and a liquid phase chip. Background technique [0002] P450 (Cytochromes P450) is the most important metabolic enzyme in the first phase of biotransformation of exogenous chemical substances in the body. Pre-carcinogens enter the body and are catalyzed by CYP450 (including epoxidation, decyclylation, oxidation, reduction, hydrolysis, etc.) Various types of chemical processes), transform into electrophilic compounds to attack intracellular biomacromolecules, and initiate carcinogenic or mutagenic processes. Cytochrome CYP450 is a group of structurally and functionally related isoenzymes encoded by superfamily genes, belonging to hemoglobin enzymes, and its subfamily CYP1A1 is closely related to smoking-induced lung cancer. [0003] CYP1A1 is an important member of the CYP450 family, locate...

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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