Amplification primer of corbiculidae shellfish mitochondria COI (Cytochrome Oxidase I) gene

A technique for amplifying primers and mitochondria, applied in the direction of DNA/RNA fragments, recombinant DNA technology, etc., can solve the problems of large inter-species variability, obstacles to the use of molecular techniques, and low amplification efficiency of COI non-specific amplification primers

Inactive Publication Date: 2014-05-21
OCEAN UNIV OF CHINA
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Problems solved by technology

[0004] Although the mtCOI sequence of Clamidae species is relatively conservative, the inter-species variability is large, which makes the amplification efficiency of COI non-specific amplification primers low, and some species cannot be amplified at all, which hinders the use of molecular techniques

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  • Amplification primer of corbiculidae shellfish mitochondria COI (Cytochrome Oxidase I) gene

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Embodiment Construction

[0007] The screening method of the amplification primer of the clam family mitochondrial COI gene of the present invention adopts three steps: a, utilize COI universal amplification primer (LCO1490F and HCO2198R, Folmer 1994) to amplify the COI sequence of part clam family species; b, from Among the part of the Clamidae COI sequences obtained above, the conserved sequence fragments were found by comparison, and multiple sets of amplification primer sequences were synthesized; c, the effect of amplifying the Clamidae mitochondrial COI gene was screened from the multiple sets of synthetic amplification primer sequences The best pair of primers.

[0008] Below in conjunction with example the present invention will be further described:

[0009] 1. Sample collection: From 2002 to 2011, a total of 126 individuals of 3 genera and 4 species of Clamidae were collected from the north and south coasts of China.

[0010] 2. DNA extraction: DNA extraction was performed on the collected s...

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Abstract

The invention relates to an amplification primer of a corbiculidae shellfish mitochondria COI (Cytochrome Oxidase I) gene. 20 corbiculidae sequences obtained by amplification of universal primers are subjected to comparative analysis by using CLUSTALW software; a plurality of pairs of primers are designed by using primer design software Primer Premier 5 in a determined conserved region; PCR (Polymerase Chain Reaction) reaction experiments are repeatedly performed by using 126 corbiculidae individuals under a 10 mu system; and finally an efficient amplification primer of a corbiculidae shellfish mitochondria COI gene sequence is sifted through detection of agarose gel electrophoresis. The amplification primer is characterized in that COIXF is 5'-TCATAAAGATATTGGGACTCT-3'; and COIXR is 5'-GGGTGACCAAAAAATCAAAAT-3'. With the adoption of the primer of the corbiculidae shellfish mitochondria COI, provided by the invention, target segments of different species of corbiculidae shellfishes can be effectively amplified, the primer has important significance to identification of corbiculidae species by using a molecule method and phylogenesis analysis, and meanwhile can provide technological conditions for researching corbiculidae systematic geography by using the COI gene and developing and protecting corbiculidae germplasm resources.

Description

technical field [0001] The invention relates to a primer for amplifying the mitochondrial COI [the cytochrome oxidase I] gene of clam shellfish. Background technique [0002] The clam family (Corbiculacea) belongs to Bivalvia, Heterodonta, and Veneroida. It is widely distributed in the waters of all continents except Antarctica. There are not many species. There are 4 genera and about 60 species in the world. species, about 5 species are known from China. It is a benthic life, mainly inhabiting the sediment bottom of rivers and lakes, and a few species inhabiting the mud bottom of mangroves. Clamidae species play an important role in maintaining the ecological balance of water areas, and some species can also be used as indicators for the detection of marine toxic pollutants tributyltin (TBT). Some species have delicious meat, high nutritional value, edible, and can also be used as medicine. They are natural bait for fish and waterfowl, and can be used as farmland fertiliz...

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/11
Inventor 于贞贞李琪孔令锋
Owner OCEAN UNIV OF CHINA
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