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Molecular marker for discriminating different ecotype populations of coilia ectenes

A molecular marker and eco-type technology, applied in the determination/inspection of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc., can solve the problems of unseen research reports and the scarcity of SINE molecular markers

Inactive Publication Date: 2014-10-15
SHANGHAI OCEAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the SINE molecular markers developed for fish are very rare, and the SINE markers provided by the present invention for the different ecotype populations of the famous and precious swordfish (Coilia nasus) have not been reported yet.

Method used

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  • Molecular marker for discriminating different ecotype populations of coilia ectenes
  • Molecular marker for discriminating different ecotype populations of coilia ectenes
  • Molecular marker for discriminating different ecotype populations of coilia ectenes

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Experimental Materials

[0025] According to the source of the samples, the anchovy is divided into 6 populations: Xiangshan Port (XS) in Zhejiang, Chongming in Shanghai (CM), Jingjiang in Jiangsu (JJ), Lake Taihu (TH), Poyang Lake in Jiangxi (PY), and Dongting Lake in Hunan (DT). The sample size of each population is 10 tails. The samples from Xiangshan Port in Zhejiang came from the aquatic product market, and the collection place is unknown, and the rest of the samples were caught on board.

[0026] Genomic DNA Extraction

[0027] The extraction of genomic DNA from the sample was performed according to the instructions of Shanghai Sangon’s UNIQ-10 Column Genomic DNA Extraction Kit. The quality and concentration of the extracted DNA were detected by an ultraviolet spectrophotometer, and stored at -20°C for future use.

[0028] site segregation

[0029] Primers were designed according to the conserved sequence of SINE, and the partial sequence of SINE in the a...

Embodiment 2

[0048] experimental method

[0049] The specific primer of SINE insertion site, its sequence is:

[0050] The upstream primer at the Locus 18 site is shown in SEQ ID NO.1,

[0051] The downstream primer at the Locus 18 site is shown in SEQ ID NO.2,

[0052] The upstream primer of the Locus 40 site is shown in SEQ ID NO.3,

[0053] The downstream primer at the Locus 40 site is shown in SEQ ID NO.4,

[0054] The upstream primer at the Locus 58 site is shown in SEQ ID NO.5,

[0055] The downstream primer at the Locus 58 site is shown in SEQ ID NO.6.

[0056] Through PCR amplification, agarose gel electrophoresis, cloning, and sequencing, the population-specific DNA fragments of the SINE locus were obtained, and different populations of the anchovy were identified according to the differences in the number and size of the amplified fragments.

Embodiment 3

[0058] experimental method

[0059] The specific primer of SINE insertion site, its sequence is:

[0060] The upstream primer at the Locus 18 site is shown in SEQ ID NO.1,

[0061] The downstream primer of the Locus 18 site is shown in SEQ ID NO.2.

[0062] Through PCR amplification, agarose gel electrophoresis, cloning, and sequencing, the population-specific DNA fragments of the SINE locus were obtained, and the sequencing results of the Locus 18 locus were analyzed, and a band with a size of about 450 bp could be amplified but not The samples with 200bp amplified bands are the anchovy from Xiangshan Port, Chongming, Jingjiang or Taihu Lake, and the samples that can amplify 450 and 200bp bands are the anchovy from Poyang Lake or Dongting Lake.

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Abstract

The invention discloses a molecular marker for discriminating different ecotype populations of coilia ectenes. A polymorphic site of coilia ectenes retrotransponsons SINE is selected as a target, and according to the flanking sequence of insertion sites, a specific primer is designed, DNA fragments with SINE inserted or with lost sites are obtained through PCR (Polymerase Chain Reaction) amplification, agarose gel electrophoresis, molecular clone and the like, and the specific DNA fragments can be used for discriminating different populations of coilia ectenes. According to the invention, the populations of coilia ectenes can be rapidly and accurately determined, and the molecular marker can be widely used in the aspects of assistant breeding, artificial releasing effect evaluation of fry, and dynamic monitoring of populations.

Description

technical field [0001] The invention relates to the technical field of molecular biology of fish, in particular, it relates to a molecular marker used for identifying different ecotype populations of the common anchovy. Background technique [0002] The definition of fish species involves two levels, one is the species level, and the other is the population level. Species identification methods include external morphological judgment method, biochemical method, and a molecular marker method developed in the past ten years. The morphological judgment method is often used for identification at the species level; for the identification at the population level, biochemical methods have been used, but this method is cumbersome and has a high rate of misjudgment, so it is basically no longer used and replaced by a molecular marker method. Because of the good stability of molecular markers, they are now widely used in theoretical studies such as fish population identification, po...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6888C12Q2600/156
Inventor 刘东朱国利唐文乔
Owner SHANGHAI OCEAN UNIV
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