Primer and probe for detecting FMO3 gene A1034T mutation of chicken

Abstract

The invention provides a primer and a probe for detecting FMO3 gene A1034T mutation of a chicken. A nucleotide sequence is shows as EQ ID NO.1-4. The primer and the probe can be utilized to quickly and sensitively detect FMO3 gene A1034T mutation sites of the chicken by adopting the real-time fluorescence quantification polymerase chain reaction (PCR) technology, namely gene type of fishlike smell sensitive genes, so that whether the chicken carries fishlike smell sensitive gene sites or not can be accurately judged. Compared with the prior art, the primer and probe have the advantages of being simple in operation, high in detection speed, capable of judging types by only one PCR, high in automation degree, high in throughout and high in sensitivity, saving time and improving detection efficiency. The whole reaction process is conducted in a closed tube, and cross pollution is reduced.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to primers and specific TaqMan MGB probes for detecting chicken FMO3 gene A1034T mutation, a detection method and a kit. Background technique [0002] In 2005, Honkatukia et al. found that a missense mutation in the flavine containing monooxygenase 3 (FMO3) gene located on chicken chromosome 8 was the cause of fishy-smelling eggs. The mutation is located on exon 7 of the FMO3 gene, and the 1034th base adenine (A) of the mutant individual cDNA sequence (GenBank accession number: AJ431390) is replaced by thymine (T), which makes the 329th base of the encoded protein The amino acid at the position is changed from threonine (T) to serine (S), so this mutation is also named T329S mutation. The mutation from threonine to serine at position 329 changes a highly conserved region (FATGY) of flavin monooxygenase. It is speculated that this region may be the binding region for substrate recogniti...

AI Technical Summary

Problems solved by technology

[0008] The purpose of the present invention is to overcome the shortcomings of the existing PCR-RFLP method to detect the existence of the mutation site, which is cumbersome, time-consuming, low sensitivity, and detection results are easily affected by experimental conditions, and to provide a method that can accurately and efficiently detect the FMO3 gene. Appropriate primers and specific TaqMan MGB probes for the A1034T mutation site, using these primers and probes, using real-time fluorescent quantitative PCR technology, can quickly and sensitively detect the genotype of the A1034T mutation site of the chicken FMO3 gene to be tested, thereby Accurately screen whether chickens carry fishy odor sensitive gene loci

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