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Specific molecular marker for identifying genetic sex of scatophagus argus as well as primers and application thereof

A molecular marker, money technology, applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc., can solve the problems of time-consuming, time-consuming and labor-intensive, high detection cost

Active Publication Date: 2020-12-04
GUANGDONG OCEAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Traditional fish sex control breeding uses test crosses to judge parental genotypes, which is time-consuming and labor-intensive. However, sex-linked molecular markers can easily and quickly realize genetic sex, which greatly saves manpower and material resources. Relevant specific molecular markers are announced, but the markers require Sanger sequencing of PCR amplification products, which is costly and time-consuming

Method used

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  • Specific molecular marker for identifying genetic sex of scatophagus argus as well as primers and application thereof
  • Specific molecular marker for identifying genetic sex of scatophagus argus as well as primers and application thereof
  • Specific molecular marker for identifying genetic sex of scatophagus argus as well as primers and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Acquisition of sex-specific markers in money fish

[0031] 1. Discovery of sex-specific molecular markers

[0032] The second-generation genome survey sequencing was performed on the DNA of the muscle tissue of 1 female and 1 male money fish, and the transcriptome sequencing of the gonad tissues of 3 female and 3 male money fish. After analyzing the sequencing data, a candidate gene for sex determination of money fish was found, dmrt1. By analyzing the resequencing data of male and female individuals, it is found that the dmrt1 gene sequence only exists in the genome of male fish and does not exist in the genome of female fish. The truncated mutant gene dmrt1b exists in both male and female fish. Compared with the gene structure of dmrt1 and dmrt1b, it is found that exon 5 of dmrt1 The similarity between the ' to 3' UTR region and the corresponding region on contig 00204 where dmrt1b is located is 78.2%. A pair of primers were designed for the gene sequence in this regi...

Embodiment 2

[0041] Application of Primer Sequences for Genetic Sex Determination of Moneyfish

[0042] Quickly and accurately identify the genetic sex of money fish, including the following steps:

[0043] (1) Collection and phenotypic sex identification of moneyfish from different geographical origins

[0044] In order to verify the accuracy of this patent for genetic sex identification, 213 goldfish (106 phenotype females and 107 phenotype males) were collected from three places in Zhanjiang, Beihai, and Zhuhai. Gender is determined by the appearance of the gonads, the ovaries of female fish are triangular, and the testes of male fish are long cystic.

[0045] (2) Genomic DNA extraction of money fish

[0046] Cut off the caudal fin of the money fish to be tested, and use the genomic DNA extraction kit to extract the money fish genome DNA step by step.

[0047] (3) Sequencing detection of the DNA of the sample to be tested

[0048] Using SEQ ID NO.3 and SEQ ID NO.4 in Example 1 to ca...

Embodiment 3

[0058] A kit for identifying the genetic sex of money fish

[0059] A kit for identifying the genetic sex of money fish, which contains primers for identifying the genetic sex of money fish; the primer sequences are shown in SEQ ID NO.3 and SEQ ID NO.4.

[0060] At the same time, the kit also contains the reagents required for PCR amplification reaction: 2×PCR mix Buffer and ddH 2 O.

[0061] The use method of the test kit comprises the following steps:

[0062] S1. Extracting the genomic DNA of the sample to be tested;

[0063] S2. Performing PCR amplification on the genomic DNA of S1 with the above primers to obtain a PCR amplification product;

[0064] S3. Perform electrophoresis on the PCR amplification product of S2. According to the electrophoresis results, if a single band is displayed in the S2 sample, it is a female fish; if two clear bands are displayed, it is a male fish.

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Abstract

The invention discloses a specific molecular marker for identifying genetic sex of scatophagus argus as well as primers and application of the specific molecular marker. The molecular marker comprisestwo DNA fragments which are partially homologous in X chromosome and Y chromosome of the scatophagus argus, a nucleotide sequence of the DNA fragment partially homologous in the X chromosome is as shown in SEQ ID NO.1, and a nucleotide sequence of the DNA fragment partially homologous in the Y chromosome is as shown in SEQ ID NO.2. The two DNA fragments partially homologous in the X chromosome and the Y chromosome of the scatophagus argus are screened out from scatophagus argus genome information, and specific primers are designed for identifying the genetic sex of the scatophagus argus, so that the genetic sex of the scatophagus argus can be quickly and accurately distinguished. The method is suitable for accurately identifying the genetic sex of the scatophagus argus in a laboratory oran aquaculture enterprise, can also be used for identifying and screening the sex of the scatophagus argus in different growth stages and different groups, and has important significance in researching a sex determination and differentiation mechanism of the scatophagus argus and controlling the sex of the scatophagus argus.

Description

technical field [0001] The invention belongs to the field of biotechnology. More specifically, it relates to a specific molecular marker for identifying the genetic sex of moneyfish, its primers and applications. Background technique [0002] Fish sex determination, differentiation mechanism and sex control technology breeding are of great significance in aquaculture research and application, and many fish are sexually dimorphic. If some fish are sexually dimorphic in growth, monosex culture can improve efficiency. Females such as Cynoglossus semilaevis (Cynoglossus semilaevis) (Chen Songlin et al., 2013) and carp (Cyprinus carpio) (Jiang et al, 2020) grow significantly faster than males, and all-feminization is one of the key technologies to improve their breeding production; Nile Fish males such as tilapia (Oreochromis niloticus) (Chen et al., 2017) and yellow catfish (Pelteobagrus fulvidraco) (Dan et al., 2018) are significantly better than females in growth, and all ma...

Claims

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Application Information

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IPC IPC(8): C12Q1/6879C12Q1/686C12N15/11
CPCC12Q1/6879C12Q1/686C12Q2600/124C12Q2565/125Y02A40/81
Inventor 黄远青江东能李广丽奥马尔·法鲁克·穆斯塔法黄洋朱春华陈华谱邓思平石红娟彭友幸
Owner GUANGDONG OCEAN UNIVERSITY
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