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A SNP molecular marker for the identification of three species of blue crabs

A technology for Scylla and Scylla simulans, which is applied in the field of SNP molecular markers, can solve the problems of damage to morphological features, inability to identify morphological features, etc., and achieve the effect of accurate identification.

Active Publication Date: 2021-09-21
NINGBO UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the above-mentioned morphological characteristics will cause damage in the process of breeding, fishing, and transportation, resulting in the inability to identify through morphological characteristics

Method used

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  • A SNP molecular marker for the identification of three species of blue crabs
  • A SNP molecular marker for the identification of three species of blue crabs
  • A SNP molecular marker for the identification of three species of blue crabs

Examples

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Effect test

Embodiment 1

[0021] Example 1: Screening of Molecular Markers to Identify Scylla

[0022] The mtDNA sequences of mud crabs, mud crabs, mud crabs, and blue crabs were obtained from the NCBI database, and the sequences were compared using AlignX (a component of Vector NTI Suite 7.1) software to find out the four kinds of mud crabs. Differential base regions in crab. The total length of the finally screened consensus sequence is 17378bp. After the analysis, it is in the 14460-14605 interval of the consensus sequence. Based on the consensus sequence, the sequences of three kinds of blue crabs, Scylla oleifera, Scylla pseudocavena, and Scylla serrata were compared , the counted SNP occurrence positions are: 14492, 14494, 14495, 14496, 14497, 14504, 14516, 14534, 14576, a total of 9 SNP sites.

[0023] The nucleotide sequence in Scylla_olivacea comprising the above-mentioned 9 SNP sites is SEQ ID NO: 1:

[0024] AAAAATTACCTAATAATGATGGTATACATACTACAACAAGCTTAAGTAAGATTCTTCGTGGTGTATCGATTCAAAAACAGGT...

Embodiment 2

[0035] Example 2 Utilize SNP sites to identify blue crabs

[0036]According to the morphological characteristics of blue crabs studied by Lin Qi et al., the used green crabs, blue crabs, and blue crabs were collected from healthy individuals in Lulin Market, Ningbo City, China, and Sanmen County, China, respectively. Take 30 individuals from each variety, take swimming foot muscle tissue respectively, use phenol-chloroform method to extract genomic DNA, dilute the extracted DNA to 100ng / microliter, and use 1.5% agarose gel electrophoresis to detect its integrity, and save it in Standby at -20°C.

[0037] 30 individuals of each species of blue crab were selected for analysis, and the PCR reaction was carried out in a Roche Light Cycle 480 instrument. The reaction system is 20 microliters, including 10 microliters LC-HRM master Mix 10 microliters, forward and reverse primers 1.5 microliters, template DNA 1.2 microliters, grade H 2 O 3 µl. The reaction program was pre-denatura...

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Abstract

The present invention provides a kind of SNP molecular marker for identifying Scylla mirabilis, Scylla pseudocavena and Scylla serrata. The nucleotide sequence of the molecular marker in Scylla serrata is SEQ ID NO: 1. The nucleotide sequence in Scylla serrata is SEQ ID NO:2, and the nucleotide sequence in Scylla serrata is SEQ ID NO:3. The present invention provides a kind of method of distinguishing Scylla serrata, Scylla pseudocaveus and Scylla serrata, described method is to judge the identity of the sample by whether there is the above-mentioned SNP molecular marker in the sample of Scylla to be detected species. The present invention has obtained the SNP molecular markers that can effectively identify the Scylla slug, Scylla pseudocavena and Scylla serrata; primers are designed in the above molecular markers, and the Scylla is typed by the high-resolution melting curve HRM, thereby rapidly , Accurately identify the germplasm of blue crabs, and provide technical support for the breeding and breeding of blue crabs.

Description

technical field [0001] The invention belongs to the technical field of molecular assisted genetic breeding, and in particular relates to a SNP molecular marker for distinguishing three kinds of blue crabs. Background technique [0002] The genus Scylla belongs to the class Crustacea, order Decapode, suborder Brachyura, and family Portunidae. Widely distributed in the coastal waters of the Indian-West Pacific Ocean, including mangrove areas and estuary bays in Southeast Asia, Australia, Japan, India, South Africa, etc. In my country, it is mainly distributed in the coastal waters of Zhejiang, Fujian, Taiwan, Guangdong, Guangxi and Hainan . The blue crab has the characteristics of large size, fast growth, strong adaptability, delicious meat, and rich nutrition. According to the trawling survey conducted by Chen Pimao and others in various reef areas from 2002 to 2004, the resource density of crabs is 180.985kg·km -2 , but the proportion of economic crabs is only 26.69%, and ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6888C12Q1/686C12N15/11
CPCC12Q1/686C12Q1/6888C12Q2600/156C12Q2527/107
Inventor 陆俊锴李荣华李永亮王春琳母昌考宋微微叶央芳刘磊史策
Owner NINGBO UNIV