Marker and method for identifying single nucleotide polymorphism of bostrichthys sinensis group

A technology of single nucleotide polymorphism and Chinese snakehead, which is applied in the fields of biochemical equipment and methods, microbial determination/inspection, etc.

Inactive Publication Date: 2011-07-27
XIAMEN UNIV
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  • Abstract
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  • Application Information

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Problems solved by technology

Among them, the populations in the South my country Sea, especially the sea area south of the Pearl River Estuary, have become the first choice for breeding seed due to their strong disease resistance, good body shape, fast growth and heat resistance, and their prices are significantly higher than those from the sea area north of the Pearl River Estuary. However, due to the

Method used

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  • Marker and method for identifying single nucleotide polymorphism of bostrichthys sinensis group
  • Marker and method for identifying single nucleotide polymorphism of bostrichthys sinensis group
  • Marker and method for identifying single nucleotide polymorphism of bostrichthys sinensis group

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Embodiment 1

[0023] 1) Select the Chinese snakeheads from Zhanjiang (south of the Pearl River estuary) and Haifeng (north of the Pearl River estuary), and use the phenol-chloroform extraction method according to the conventional method (see "Molecular Cloning Experiments" translated by Huang Peitang et al. Guide" (Third Edition), Science Press, September 2002) to extract the DNA of S. sinensis;

[0024] 2) Design and synthesize a pair of selective amplification primers

[0025] tRNA-proH: 5'-gtgtgagaag ggagattcta actcccaacc-3'

[0026] tRNA-proL: 5'-acgggatggt ggttcgtggt at-3';

[0027] Carry out two PCR amplification reactions on the DNA of Channa sinensis:

[0028] The first PCR amplification, that is, the positive detection reaction PCR amplification program was: pre-denaturation at 95°C for 5 min; each cycle of denaturation at 95°C for 30 s, annealing at 58°C for 30 s, and extension at 72°C for 30 s, for a total of 35 cycles.

[0029] The second PCR amplification, that is, the selec...

Embodiment 2

[0033] Similar to Example 1, the difference is that the Chinese snakehead from the south of the Pearl River estuary was taken from Zhuhai; the sample from the north of the Pearl River estuary was selected from Xiamen. When electrophoresis test results, under the premise of ensuring that the positive test reaction is normal (that is, there is a positive amplification product), in the selective PCR amplification, if there is an amplification product of 330bp, the DNA sample is from Xiamen. If there is no amplification product, the DNA sample is from Zhuhai (see figure 1 and 2 ).

Embodiment 3

[0035] Similar to Example 1, the difference is that the Snakena sinensis in the south of the Pearl River estuary was taken from the North Sea; the sample in the north of the Pearl River estuary was taken from Ningde. When electrophoresis test results, under the premise of ensuring that the positive test reaction is normal (that is, there is a positive amplification product), in the selective PCR amplification, if there is an amplification product of 330bp, the DNA sample is from Ningde, if there is no amplification product, the DNA sample comes from the North Sea (see figure 1 and 2 ).

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Abstract

The invention discloses a marker and method for identifying single nucleotide polymorphism of a bostrichthys sinensis group and relates to a method for identifying a bostrichthys sinensis group. A single nucleotide polymorphism marker of the bostrichthys sinensis group is positioned on a 29th basic group at a 5' end of a praline tRNA gene in a bostrichthys sinensis mitochondrial genome. The method for identifying single nucleotide polymorphism of the bostrichthys sinensis group comprises the steps of: extracting a genome DNA of a bostrichthys sinensis; carrying out PCR (Polymerase Chain Reaction) amplification on the praline tRNA gene in the bostrichthys sinensis mitochondrial genome by using the DNA as a template, carrying out electrophoretic analysis on the PCR amplification products, determining that the sample is from the bostrichthys sinensis group in North of Pearl River Estuary when 330-bp PCR amplification products are tested, and determining that the sample is from the bostrichthys sinensis group in South of Pearl River Estuary when 330-bp PCR amplification products are not tested. The method for identifying single nucleotide polymorphism of the bostrichthys sinensis group is simple and easy, accurate and fast, impartial and subjective as well as economical and practical.

Description

technical field [0001] The invention relates to a method for identifying a population of Channa sinensis, in particular to a single nucleotide polymorphism marker and a method for identifying a population of Channa sinensis. Background technique [0002] Chinese snakehead (Bostrychus sinensis), also known as mullet, salmon, bamboo shoot shell fish, earth fish, crab tiger, belongs to Perciformes (Perciformes), Gobioidei (Gobioidei), pond snakehead family (Eleotridae) , Wutang Channa (Bostrychus), whose wild populations are widely distributed in the intertidal sea area from the south of the Yangtze River Estuary to Southeast Asia, is a rare and valuable seafood in the southeast coast of my country. At present, the market price can reach 160 yuan / kg (Zhang Jiandong , the growth, growth model and life history types of Channa sinensis. Chinese Journal of Ecology [J], 200222(6): 841-846). At present, Channa sinensis is cultivated in Guangdong, Guangxi, Fujian, and Zhejiang, but the...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 丁少雄王航俊张丽艳王军
Owner XIAMEN UNIV
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