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A sex-specific molecular marker primer, identification method and application of pomfret ovata

A technique for oval pomfret and molecular markers, which is applied in biochemical equipment and methods, microbiological measurement/inspection, DNA/RNA fragments, etc., can solve the problems of oval pomfret death and inability to identify the sex of oval pomfret , to achieve the effects of improving breeding efficiency, avoiding inbreeding depression, and high identification success rate

Active Publication Date: 2022-06-21
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) Usually in production practice, it is impossible to identify the sex of pompano ovata by phenotype
[0005] (2) In scientific research, the physiological sex of the pomfret can be identified by means of shape observation and tissue section, but the gonad tissue needs to be dissected and collected, resulting in the death of the pomfret
[0006] (3) At present, there is no technology for live sex identification by means of biotechnology

Method used

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  • A sex-specific molecular marker primer, identification method and application of pomfret ovata
  • A sex-specific molecular marker primer, identification method and application of pomfret ovata
  • A sex-specific molecular marker primer, identification method and application of pomfret ovata

Examples

Experimental program
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Embodiment 1

[0032] The oval pomfret sex-specific molecular marker primer provided in this example includes a forward primer and a reverse primer, wherein the base sequence of the forward primer is shown in SEQ ID NO: 1, and the base sequence of the reverse primer is shown in SEQ ID NO: 1. The sequence is shown in SEQ ID NO:2.

[0033] specific:

[0034] The forward primer was 5'-ctccaacagctgcccaaac-3'.

[0035] The reverse primer was 5'-ccacgatccgcacttacaac-3'.

[0036] The primer design is based on the following principles:

[0037] A full-sib family of ovoid pomfret was collected, including 2 parents and 100 offspring individuals, and the physiological sex was identified after autopsy. High-throughput sequencing was used to develop markers for parents and progeny, using the genome sequence of the oval pomfret as the reference sequence, and linkage analysis was used to locate the sex quantitative trait loci and locate them in a single interval. Significant sex-linked indels within th...

Embodiment 2

[0057] The method for identifying males and females of the oval pomfret provided in this embodiment includes the following steps: extracting the genomic DNA of the oval pomfret to be tested, using the primers in Example 1 to perform PCR amplification, and subjecting the amplified product to capillary electrophoresis detection amplification Addition sequences, in which females are homozygous for a 10bp insertion and males are homozygous or heterozygous for a 10bp deletion.

[0058] Specifically include the following steps:

[0059] (1) DNA extraction

[0060] 112 oval pomfrets (unknown male and female) to be tested were selected, about 1 square centimeter of fin rays were cut from the living body, and stored in 95% alcohol. After sampling, the oval pomfrets were put back into place. Use DNA extraction kits or phenol imitation method to extract tissue DNA.

[0061] (2) Amplification of sex-specific markers of oval pomfret

[0062] The forward primer in Example 1 was 5'-ctccaa...

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Abstract

The invention discloses a sex-specific molecular marker primer for ovoid pomfret. The primer includes a forward primer and a reverse primer. The base sequence of the forward primer is shown in SEQ ID NO: 1, and the reverse primer The base sequence of the primer is shown in SEQ ID NO:2. Also disclosed is a method for using the above primers to identify the male and female of the ovoid pomfret and the application of the above primer or the method in preparing a kit or biological preparation for identifying the male and female of the ovate pomfret. The primer has good specificity, the identification method is convenient to operate, simple and easy to implement, the identification success rate is high, and there is no harm to the living pomfret, and the sex of the pomfret can be quickly and accurately identified in batches at one time, which is of great importance. Scientific research value and practical application value.

Description

technical field [0001] The invention belongs to the technical field of genetics and breeding of marine economic fish, in particular to a sex-specific molecular marker primer of oval pomfret, an identification method and application thereof. Background technique [0002] Oval pomfret (Trachinotus ovatus), belonging to the order Perciformes, Trevidae, Pomfret genus, commonly known as golden pomfret, yellow pomfret, and yellow pomfret, is a high-quality breeding object developed in China in the past ten years. The best breed for breeding, the current domestic output of breeding has exceeded 100,000 tons. There is no morphological difference between male and female individuals in the early embryonic development and juvenile stage. It is impossible to identify the sex of male and female from the body size and genital characteristics. Sex can only be determined by distinguishing male and female reproductive organs after dissection. identification. Gender identification is the mo...

Claims

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Application Information

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