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Specific molecular marker, primer and method for identifying genetic sex of selenotoca multifasciata and application of specific molecular marker and primer

A technology of molecular markers and identification methods, applied in the direction of biochemical equipment and methods, measurement/testing of microorganisms, DNA/RNA fragments, etc., to achieve good application value

Pending Publication Date: 2021-11-30
GUANGDONG OCEAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, no specific molecular markers related to genetic sex have been found in C. multistripe, so the development and screening of specific molecular markers for C. multistripe play an important role in sex-controlled breeding. Fish genetic sex identification and parthenoculture technology have good application value

Method used

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  • Specific molecular marker, primer and method for identifying genetic sex of selenotoca multifasciata and application of specific molecular marker and primer
  • Specific molecular marker, primer and method for identifying genetic sex of selenotoca multifasciata and application of specific molecular marker and primer
  • Specific molecular marker, primer and method for identifying genetic sex of selenotoca multifasciata and application of specific molecular marker and primer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1 Accessible for gender-specific markers of multiped money butterflyfish

[0038] 1. Development of gender-specific molecular markers

[0039] The inventor's topic group uses a tail male multi-line butterfly fish Illumina genome sequencing data, and the candidate gene DMRT1 is compared on a multi-straightened butterflyfish through the same-grade money fish (SCATOPHAGUS ARGUS). Butterfly DMRT1 gene sequence. By alignment analysis, the separated multi-line butterflyfish DMRT1 partial genome sequence is found that DMRT1 has another copy of DMRT1 in addition to normal DMRT1, and has obtained part of its sequence. According to the sequence comparison analysis of multiped money butterfly DMRT1 and its homologous genes, it is designed to verify whether or not to chain with gender chains.

[0040] Design a pair of primers for DMRT1 and its homologous gene sequence, as shown in SEQ ID NO: 3 ~ 4:

[0041] Positive primer SEQ ID NO.3: 5'-agagctTgttctctGataActtaAg-3 ';

[0042] ...

Embodiment 2

[0049] Example 2 Application of multiped money butterflyfish genetic gender identification primer sequence

[0050] 1. Quick identification multi-striped money butterflyfish genetic gender

[0051] (1) Collection and phenotype gender identification of multiped money butterflyfish

[0052] Collect 84 tail multi-line-torn butterflyfish purchased from Zhanjiang Water Products Wholesale Market (32 Tail Table Female and 52 Tail Mouth), the gender of multiped money butterflyfish is determined by the gland tissue slices, female Fish ovary has an oocyte at which II, III or IV, and the male fish briquette has racemic cells at all levels.

[0053] (2) Multipined money butterflyfish genome DNA extraction

[0054] Cut the tapered fins of the multi-straw-to-white fish, using genomic DNA extraction kits (marine animal tissue genomic extract kit, Guangzhou Dongsheng Biotechnology Co., Ltd., Item N1173), and extract multi-straighten money butterfly genome DNA by step .

[0055] (3) Sequencing det...

Embodiment 3

[0062] Example 3 A quick identification of multi-striped money butterflyfish genetic sex kit

[0063] A kit for identifying a multi-striped money butterflyfish genetic sex, the kit containing a primer for identifying the genetic sex of multi-striped money butterflyfish, the primer sequence as follows:

[0064] Upstream primer F: 5'-agagctTgtttctctGataActtaAg-3 '(SEQ ID NO: 3)

[0065] Downstream primers R: 5'-gtcaccatcgatgatgagct-3 '(SEQ ID NO: 4)

[0066] At the same time, the kit also contains reagents required for PCR amplification reactions: 2 × PCR Mix Buffer and DDH 2 O.

[0067] The method of use of the kit includes the steps of:

[0068] S1. Extract multi-striped money squid sample genome DNA;

[0069] S2. PCR amplification is performed by the genomic DNA of S1 in the above primer;

[0070] S3. The PCR amplification product of S2 is electrophoresed according to the electrophoresis results. If the S2 sample is displayed, it is a female fish; if the two strips are displayed,...

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Abstract

The invention discloses a specific molecular marker for genetic sex of selenotoca multifasciata. The molecular marker comprises two DNA fragments which are partially homologous in X chromosome and Y chromosome of the selenotoca multifasciata, and the nucleotide sequences of the DNA fragments are shown as SEQ ID NO: 1-2 in sequence. According to the invention, the two homologous DNA fragments on the X chromosome and the Y chromosome of the selenotoca multifasciata are screened from genome information of the selenotoca multifasciata for the first time, and the method for identifying the genetic sex of the selenotoca multifasciata is established, so that the genetic sex of the selenotoca multifasciata can be quickly and accurately distinguished. The detection and identification method provided by the invention can be used for identifying and screening the sex of the selenotoca multifasciata, and is suitable for accurately identifying the genetic sex of the selenotoca multifasciata in laboratories or aquaculture enterprises. The method is of great significance in studying sex determination and differentiation mechanisms of the selenotoca multifasciata and realizing sex control of the selenotoca multifasciata.

Description

Technical field [0001] The invention belongs to the field of biotechnology. More specifically, a specific molecular marker, primer, primer, primer, primer, primer, primer, primers, identification of multi-striped money butterflyfish. Background technique [0002] Silenotoca Multifasciata, also known as Silver Miyu, is affiliated with percifores, Money Fish (Scatophagidae), Qian Butterfly Fish, the native of Southeast Asia, Philippines, Indonesia and other countries coastal areas It also has a distribution in the East my country Sea and South China Sea. It has been broken through artificial breeding has been technically, and it is an omitcological fish, and excellent in farming. It is an important economic fish that is equally ornamental and edible. It can bring huge economic benefits, social benefits and ecological benefits. [0003] Since many farmers have a gender bismuth, there is a significant gender difference in the morphological structure of maid and male, and gender contr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12Q1/6879C12Q1/686C12N15/11
CPCC12Q1/6888C12Q1/6879C12Q1/686C12Q2565/125
Inventor 江东能黄远青李广丽邓秋敏彭友幸黄洋朱春华
Owner GUANGDONG OCEAN UNIVERSITY
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