Trachinotus ovatus gender specific molecular marker primer and application thereof

An oval pomfret, specific technology, applied in the direction of microbial determination/inspection, biochemical equipment and methods, DNA/RNA fragments, etc., can solve the problems of anatomical individual death, inefficiency, and inability to achieve large-scale identification, etc.

Active Publication Date: 2020-07-07
SOUTH CHINA SEA FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] (1) In scientific research work, the biological sex of pomfret can be identified through dissection and other methods, but dissection will inevitably lead to the death of the dissected individual, and this method is inefficient and cannot achieve large-scale identification
[0005] (2) In production practice, it is impossible to identify the sex of live pomfrets by phenotype
[0006] (3) There is currently no technology for live sex identification through biotechnological means

Method used

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  • Trachinotus ovatus gender specific molecular marker primer and application thereof
  • Trachinotus ovatus gender specific molecular marker primer and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0030] The sex-specific molecular marker primers for ovoid pomfret provided in this example, the primers are primers P2, and the primers P2 include forward primers P2-F and reverse primers P2-R, and the forward primers P2-F The base sequence of the reverse primer P2-R is shown in SEQ ID NO:1, and the base sequence of the reverse primer P2-R is shown in SEQ ID NO:2.

[0031] Specifically, the base sequence of primer P2 is as follows:

[0032] Forward primer P2-F: 5'-CATGGACAAGAAGGTGGTGC-3';

[0033] Reverse primer P2-R: 5'-TACCCAGTGCAAGCTCTCTC-3'.

[0034] A full-sib family of Pompano ovata was collected, including 2 parents and 100 offspring individuals, and the physiological sex was identified after dissection. High-throughput sequencing was used to develop markers for parents and offspring using the genome sequence of the pomfret ovata as a reference sequence, and linkage analysis was used to locate the sex quantitative trait loci and locate them in a single interval. Sig...

Embodiment 2

[0040] The method for male and female identification of ovoid pomfret provided by this embodiment comprises the following steps: using PCR amplification to amplify specific fragments in the genomic DNA of male and female individuals, specifically including: designing the primers in Example 1, according to the embodiment The method in 1 extracts the genomic DNA of the ovoid pomfret to be tested, adopts the primers in Example 1, and uses the extracted genomic DNA as a template to carry out PCR amplification, using gel electrophoresis (specifically, an automatic sequencer ABI 3730XL can be used) The amplified products were detected, and male and female individuals were identified according to the electrophoresis results.

[0041] The implementation object of the present embodiment is oval pomfret, gets 47 females and 47 males, sex is identified by anatomy, and then the feasibility of this primer is verified.

[0042] (1) Extraction of DNA

[0043] Cut about 1 square centimeter o...

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Abstract

The invention discloses a trachinotus ovatus gender specific molecular marker primer. The primer is a primer P2 which comprises a forward primer P2-F and a reverse primer P2-R; and the base sequencesof the forward primer P2-F and the reverse primer P2-R are respectively as shown in SEQ ID NO: 1 and SEQ ID NO: 2. The invention also discloses a test kit used for gender determination of trachinotusovatus, and an application of the above-mentioned primer or the test kit in gender determination of the trachinotus ovatus. The primer provided by the invention is good in specificity, high in sensitivity and high in accuracy, does not harm trachinotus ovatus living bodies, can quickly and accurately identify the gender of the trachinotus ovatus in batches at a time, and has important scientific research value and practical application value.

Description

technical field [0001] The invention belongs to the technical field of sex of pomfret ovata, in particular to a sex-specific molecular marker primer and application thereof. Background technique [0002] Oval pomfret (Trachinotus ovatus), belonging to Perciformes, Trevalidae, Pomfret genus, commonly known as golden pomfret, yellow pomfret, yellow waxed pomfret. After the breakthrough of artificial induced spawning in 2000, large-scale aquaculture began in China. At present, it has developed into an important marine aquaculture species in my country, with an output of more than 100,000 tons. This species has the characteristics of rapid growth, strong disease resistance, and turbulent flow. The best species for large-scale farming in deep and open seas in the future. Although the breeding scale has increased, the breeding work has lagged behind. The important limiting factor is that the pomfret can't distinguish the sex from the appearance, which makes it impossible to contro...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6879C12Q1/6888C12Q1/686C12N15/11
CPCC12Q1/686C12Q1/6879C12Q1/6888C12Q2565/125
Inventor 张殿昌郭梁杨静文刘宝锁郭华阳朱克诚张楠杨权
Owner SOUTH CHINA SEA FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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