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Trachinotus ovatus sex specific molecular marker primer, identification method and application of primer and method

A technology 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: 2020-05-01
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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  • Trachinotus ovatus sex specific molecular marker primer, identification method and application of primer and method
  • Trachinotus ovatus sex specific molecular marker primer, identification method and application of primer and method
  • Trachinotus ovatus sex specific molecular marker primer, identification method and application of primer and method

Examples

Experimental program
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Effect test

Embodiment 1

[0032] The sex-specific molecular marker primer for ovoid pomfret 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 The sequence is shown in SEQ ID NO:2.

[0033] specific:

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

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

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

[0037] 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 significantly linked to sex in inter...

Embodiment 2

[0057] The method for male and female discrimination of the ovoid pomfret provided in this example comprises the following steps: extracting the genomic DNA of the ovate pomfret to be tested, performing PCR amplification using the primers in Example 1, and performing capillary electrophoresis detection amplification on the amplified product. Augmentation sequence, in which the female is homozygous for a 10bp insertion, and the male is homozygous or heterozygous for a 10bp deletion.

[0058] Specifically include the following steps:

[0059] (1) Extraction of DNA

[0060] Select 112 ovate pomfrets (unknown male and female) to be tested, cut about 1 square centimeter of fin rays from the living body, store them in 95% alcohol, and return the ovate pomfret after sampling. DNA extraction kit or phenol imitation method was used to extract tissue DNA.

[0061] (2) Amplification of allospecific markers in pompano ovata

[0062] The forward primer in Example 1 is 5'-ctccaacagctgccc...

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Abstract

The invention discloses a trachinotus ovatus sex specific molecular marker primer. The primer comprises a forward primer and a reverse primer, wherein a base sequence of the forward primer is as shownin SEQ ID NO: 1, and a base sequence of the reverse primer is as shown in SEQ ID NO: 2. The invention also discloses a method for identifying male and female of trachinotus ovatus by using the aboveprimer, and an application of the above primer or method in preparing a kit or biological preparation for identifying male and female of trachinotus ovatus. The primer has good specificity, and the identification method is convenient to operate, is simple and easy to implement, is high in identification success rate, does not harm the live body of trachinotus ovatus, can quickly and accurately identify the sex of trachinotus ovatus in batches at once, and has important scientific research value and practical application value.

Description

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

Claims

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Application Information

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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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