Specific molecular marker primer for genders of trachinotus ovatus, kit and application of kit

A technology of oval pomfret and molecular markers, which is applied in biochemical equipment and methods, microbiological measurement/inspection, DNA/RNA fragments, etc., can solve the problem of inability to identify the sex of oval pomfret live bodies, the inability to realize large-scale identification, Anatomy of individual death and other issues to achieve the effect of avoiding inbreeding depression, breaking through technical bottlenecks, and high accuracy

Active Publication Date: 2020-05-29
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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  • Specific molecular marker primer for genders of trachinotus ovatus, kit and application of kit
  • Specific molecular marker primer for genders of trachinotus ovatus, kit and application of kit

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 are primers P1, and the primers P1 include forward primers P1-F and reverse primers P1-R, and the forward primers P1-F The base sequence of the reverse primer P1-R is shown in SEQ ID NO:1, and the base sequence of the reverse primer P1-R is shown in SEQ ID NO:2.

[0031] Specifically, the base sequence of primer P1 is as follows:

[0032] Forward primer P1-F: 5'-AACTCTATGCACCGACCTGA-3';

[0033] Reverse primer P1-R: 5'-CATGAACAGACACGAGCACC-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 mark the parents and offspring with the genome sequence of the pomfret pomfret as a reference sequence, and linkage analysis was used to locate the sex quantitative trait loci and locate them in a single interval. Development of indels sign...

Embodiment 2

[0040] The method for male and female identification of oval 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, and adopts gel electrophoresis (specifically using an automatic sequencer ABI 3730XL) to detect The amplified products were identified as male and female individuals according to electrophoresis results.

[0041] The object of implementation in this embodiment is pomfret ovata, and 47 females and 47 males were taken, and the sexes were identified through anatomy, and then the feasibility of the primer was verified.

[0042] (1) Extraction of DNA

[0043] Cut about 1 square c...

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Abstract

The invention discloses a specific molecular marker primer for genders of trachinotus ovatus. The specific molecular marker primer is a primer P1 including a forward primer P1-F and a backward primerP1-R, and base sequences of the forward primer P1-F and a backward primer P1-R are respectively represented by SEQ ID NO:1 and SEQ ID NO:2. The invention further discloses a kit for identifying the genders of the trachinotus ovatus and application of the primer or the kit in identification of the genders of the trachinotus ovatus. The specific molecular marker primer is good in specificity and high in sensitivity and accuracy, is harmless to living trachinotus ovatus, can be used for rapidly and accurately identifying the genders of the trachinotus ovatus in batch at a time and has important scientific research values and practical application values.

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, kit and application of pomfret ovata. Background technique [0002] Oval pomfret (Trachinotus ovatus), belongs 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 my country. At present, it has developed into an important marine aquaculture species in China, 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 scale of its breeding has increased, the breeding work has lagged behind. The important limiting factor is that the oval pomfret can't distinguish the sex from its appearance, which makes...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6879C12Q1/686C12N15/11
CPCC12Q1/686C12Q1/6879C12Q2565/125
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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