Paralichthys olivaceus fertility related SNP molecular marker and screening method and application thereof

Abstract

The invention discloses a paralichthys olivaceus fertility related SNP molecular marker and a screening method and application thereof. The screening method comprises the following steps: separately carrying out cyp21a gene re-sequencing on fertile paralichthys olivaceus and infertile DH paralichthys olivaceus at first to preliminarily screen difference SNP sites; verifying the SNP sites on a large scale by using a fertile population capable of spawning normally in a breeding season and DH infertile population incapable of spawning; carrying out parting on the SNP sites, which are obtained instep (1), of the paralichthys olivaceus by using a multiple SNaPshot SNP parting method; and carrying out statistics on genotype frequency of the SNP sites which are successful in parting by using SPSS software, and carrying out difference significance analysis by using chi-square test to screen out SNP sites related to paralichthys olivaceus fertility. In the invention, a paralichthys olivaceus fertility research model is established, a method for rapidly establishing a large number of 21OHD animal models is provided, and SNP sites of fertility related cyp21a genes are obtained by screening.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a cyp21a gene SNP molecular marker, screening method and application related to the fertility of a flounder model of 21OHD (21 hydroxylase deficiency). Background technique [0002] cyp21a locates in human 6p21.3 and consists of two genes, true gene and pseudogene. cyp21a is a functional gene encoding 21 hydroxylase, its mutation can lead to the structural change of 21 hydroxylase and the loss or decrease of activity, thus producing a negative feedback effect to increase the synthesis of CRH and ACTH, leading to adrenal hyperplasia, clinically manifested as female Symptoms of virilization, infertility (CAH). At present, in order to study the CAH caused by cyp21a mutation, that is, 21OHD, people have constructed a naturally mutated 21OHD mouse model and a zebrafish model after cyp21a knockout, but the mouse model is difficult to feed, and the zebrafish model can only show cyp21a Kno...

AI Technical Summary

Problems solved by technology

At present, in order to study the CAH caused by cyp21a mutation, that is, 21OHD, people have constructed a naturally mutated 21OHD mouse model and a zebrafish model after cyp21a knockout, but the mouse model is difficult to feed, and the zebrafish model can only show cyp21a The problem with knocking out this mutant

The etiology of human 21OHD is complex, usually including homozygous mutations and complex heterozygous mutations, among which the most common homozygous mutations are Val281Leu mutations, followed by Pro30Leu, Arg339His, and Pro453Ser mutations, which may lead to a 30%-60% decrease in enzyme activity

[0003] Therefore, the current experimental animal models related to human fertility are difficult to meet the research needs; therefore, it is necessary to construct an experimental model that can meet the requirements of human fertility.

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