Detection method of halothane genotype in swine

A detection method and porcine fluorocarbon technology, applied in the field of bioengineering, can solve the problems of toxic detection reagents, many operation steps, complicated processes, etc., and achieve the effects of fast reaction, pollution avoidance, and steps reduction.

Inactive Publication Date: 2009-02-04
SHANGHAI ANIMAL EPIDEMIC PREVENTION & CONTROL CENT
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Problems solved by technology

[0003] The technical problem to be solved by the present invention is to provide a method for detecting the genotype of the porcine halothane gene. The method for detecting the genotype of the porcine halothane gene should solve the problem that the method for detecting the genotype of the porcine halothane gene in the prior art is accurate. Technical problems such as low rate, many operation steps, complicated process, long time, and toxic detection reagents

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  • Detection method of halothane genotype in swine
  • Detection method of halothane genotype in swine
  • Detection method of halothane genotype in swine

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Embodiment Construction

[0022] Detection method of the present invention comprises the steps:

[0023] 1. Primer Design

[0024] By performing base sequencing on the porcine halothane gene base fragment amplified by ordinary PCR (as shown in SEQID NO: 1), it is determined that the mutation site of the porcine halothane gene is the 494th cytosine (C) mutation is Thymine (T), two sets of primers were designed according to the nucleotide sequences at both ends of the mutation site, in which the 3' end of the upstream primer of one set matched the gene site of the wild type, and the 3' end of the other set of upstream primers matched the gene site of the mutant type The gene loci matched; the downstream primers of the two groups were the same.

[0025] The primer sequences are:

[0026] Wild-type upstream primer: 5′CAA TGG TGT GGC CGT GT 3′

[0027] Wild-type downstream primer: 5′GCC CAG ACC TGG TGA CAT A 3′

[0028] Mutant upstream primer: 5′CAA TGG TGT GGC CGT GC 3′

[0029] Mutant downstream prim...

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Abstract

The invention provides a genotype test method for a pig halothane gene. Two groups of different primers are schemed out according to the base sequences of two ends of the mutant site of the pig halothane gene, sample DNA respectively receives the PCR amplification with simultaneously, the change of the product amount is monitored through the fluorescent signal intensity change to judge the genotype of the sample DNA. The invention solves the technical problems of low accuracy, many operation steps, complicate process, long time and toxic test reagents of the methods for testing the genotype of the pig halothane gene in the prior art, improves the test accuracy, simplifies the operation procedures, and avoids the use of the toxic and harmful reagents.

Description

Technical field: [0001] The invention belongs to the field of bioengineering, and in particular relates to a gene detection method, in particular to a method for detecting porcine halothane gene genotypes, in particular to a method for detecting different genotypes of porcine halothane gene by using fluorescent quantitative PCR technology. Background technique: [0002] Porcine halothane gene, also known as skeletal muscle calcium ion release channel gene or ryanodine receptor gene, consists of a pair of alleles Hal N and Hal n Composed, and constituted Hal NN (wild type), Hal Nn (heterozygous) and Hal nn (Mutant type) 3 genotypes, which are a main gene that causes porcine stress syndrome. The typical symptoms of porcine stress syndrome are malignant hyperthermia, muscle rigidity, sudden death, and poor meat quality after slaughtering, resulting in poor quality Meat. Pig stress death and poor quality meat caused by porcine stress syndrome have caused huge losses to pig ...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11G01N21/64
Inventor 李建颖刘炜吴昊旻马芳芳黄国庆
Owner SHANGHAI ANIMAL EPIDEMIC PREVENTION & CONTROL CENT
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