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Specific molecular marker and primer for accurately identifying heredity of scatophagus argus

A molecular marker, money fish technology, applied in the determination/inspection of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc., can solve the problems of no specific molecular markers, time-consuming and labor-intensive

Active Publication Date: 2018-06-22
GUANGDONG OCEAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Traditional fish sex control breeding requires test crossing to determine whether the parental genotype is XX pseudo-male fish, which is time-consuming and labor-intensive
[0004] However, sex-linked molecular markers can easily and quickly achieve hereditary sex, which greatly saves manpower and material resources. However, no specific molecular markers related to hereditary sex have been found in money fish.

Method used

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  • Specific molecular marker and primer for accurately identifying heredity of scatophagus argus
  • Specific molecular marker and primer for accurately identifying heredity of scatophagus argus
  • Specific molecular marker and primer for accurately identifying heredity of scatophagus argus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1 Obtaining sex-specific markers of money fish

[0045] 1. Extraction of gonad RNA

[0046] Three female goldfishes were taken, and the male and female gonads were taken separately, frozen in liquid nitrogen, and then transferred to a -80°C refrigerator for extraction of total RNA. The total RNA of the sample was extracted by RNAiso, and then detected by 1% agarose gel electrophoresis. Two bands of 28S and 18S were clearly visible. The OD value and Concentration, OD260 / 280 values ​​are between 1.95 and 2.00. The total RNA samples were sent to Biosequencing Company on dry ice for transcript assembly and sequencing analysis.

[0047] 2. Gonad transcriptome sequencing assembly and annotation

[0048] After obtaining the raw data, the Raw reads are filtered and assembled by the De novo assembly software Trinity, and the assembled sequences are deredundant and spliced ​​by the software TGICL to obtain the longest possible non-redundant Unigene set, and Unigene set ...

Embodiment 3

[0070] Example 3 A kit for rapidly identifying the genetic sex of money fish

[0071] A kit for identifying the genetic sex of money fish, said kit containing primers for identifying the genetic sex of money fish; said primer sequence is as follows:

[0072] Upstream primer F: 5′-ACGAGAGCCTGGAGAGCCTCAT-3′ (SEQ ID NO: 3)

[0073] Downstream primer R: 5′-TGCGGCACCACTGTGTAACTGA-3′ (SEQ ID NO: 4)

[0074] At the same time, the kit also contains the reagents required for PCR amplification reaction: 2× PCR mix Buffer and ddH 2 O.

[0075] The using method of described test kit comprises the steps:

[0076] S1. Extracting the genomic DNA of the sample to be tested;

[0077] S2. Perform PCR amplification on the genomic DNA of S1 with the above primers;

[0078] S3. Perform electrophoresis on the PCR amplification product of S2 to obtain a single electrophoresis band, cut it off, and purify;

[0079] S4. Sequence the PCR purified product of S3, the sequencing primer is the upstre...

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Abstract

The invention discloses a specific molecular marker for the heredity of scatophagus argus. The molecular marker comprises two DNA fragments which are homoeologous in the X chromosome and the Y chromosome of scatophagus argus, the nucleotide sequence of the DNA fragment on the X chromosome is shown as SEQ ID NO: 1, and the nucleotide sequence of the DNA fragment on the Y chromosome is shown as SEQID NO: 2. The two DNA fragments which are homoeologous in the X chromosome and the Y chromosome of scatophagus argus are screened from scatophagus argus genome information for the first time, a specific primer is designed to identify the heredity of scatophagus argus, and the genetic genders of scatophagus argus can be quickly and accurately distinguished. The method is suitable for laboratories or aquaculture enterprises to accurately identify the genetic genders of scatophagus argus. The method for detecting the genetic genders of scatophagus argus can be used for gender identification and screening of different scatophagus argus groups in different growth stages, and plays an important role in researching gender determination and differentiation mechanism of scatophagus argus and achieving gender control of scatophagus argus.

Description

technical field [0001] The invention belongs to the field of biotechnology. More specifically, it relates to a specific molecular marker and primers for accurately identifying the genetic sex of money fish. Background technique [0002] The sex determination, differentiation mechanism and sex control technology breeding of fish are of great significance in aquaculture research and application. Many fish are sexually dimorphic. Cynoglossus semilaevis ),flounder( Hippoglossus hippoglossus L.) and carp ( Cyprinus carpio ) and other females grow significantly faster than males. Realizing all-feminization breeding through sex control technology is one of the key technologies to improve its breeding production (Chen Songlin et al., 2013; Shao et al., 2014); Nile tilapia ( Oreochromis niloticus ) and yellow catfish ( Pelteobagrus fulvidraco ) and other fish males are significantly better than females in growth, and it is expected to achieve all-masculinization in production...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6879C12N15/11
CPCC12Q1/6879
Inventor 江东能李广丽奥马尔·法鲁克·穆斯塔法朱春华陈华谱邓思平吴天利梁志华向玲杨尉古皓天
Owner GUANGDONG OCEAN UNIVERSITY
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