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Amplification primer of tridacnidae shellfish mitochondria COI (cytochrome oxidase I) gene

A gene amplification and mitochondrial technology, applied in the direction of DNA / RNA fragments, recombinant DNA technology, etc., can solve the problems of low amplification efficiency, large variation, primers hinder molecular identification research, etc., and achieve the effect of efficient amplification

Inactive Publication Date: 2012-12-12
OCEAN UNIV OF CHINA
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Problems solved by technology

[0004] However, the mutation rate of shellfish mtCOI sequence is significantly higher than that of other groups of animals, and the mtCOI sequence variation among different mollusk species, even related species or even different groups within a species, is very large, which led to Folmer et al. 1994 The amplification efficiency of the COI universal primers LC01490 and HCO2198 designed in 2009 was extremely low in this animal group
The lack of primers with high amplification efficiency has seriously hindered the application of this molecular marker to the molecular identification of bivalve molluscs

Method used

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Embodiment Construction

[0007] The screening method of the Tridacnaceae shellfish mitochondrial COI gene amplification primer of the present invention adopts four steps: a, on the National Center for Biotechnology Information of the United States (NCBI, http: / / www.ncbi.nlm.nih.gov / ) Download the existing Tridacnaceae sequence; b, use the COI universal amplification primers (LCO1490F and HCO2198R, Folmer 1994) to amplify the COI sequences of some Tridacnaceae species; c, from the Tridacnaceae mtCOI sequence obtained above, Find the conserved sequence fragments by comparison, and synthesize multiple sets of amplification primer sequences; d, screen out a pair of primers with the best effect of amplifying the Tridacaceae mitochondrial COI gene from the multiple sets of synthetic amplification primer sequences.

[0008] The present invention will be further described below in conjunction with embodiment:

[0009] 1. Download 52 COI sequences of 5 species of Tridacnaceae molluscs from the National Center ...

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Abstract

The invention relates to an amplification primer of a tridacnidae shellfish mitochondria COI (cytochrome oxidase I) gene. The known 52 tridacnidae sequences in NCBI (National Center for Biotechnology Information) are utilized, two tridacnidae sequences obtained by amplifying a universal primer are jointly utilized, a CLUSTALW software is used for comparison analysis, a plurality of pairs of primers are designed by using a primer design software Primer Premier 5 in a determined conversation region, 43 tridacnidae individuals are repeatedly subjected to PCR (Polymerase Chain Reaction) experiments in a 10 mul system and are finally detected through agarose gel electrophoresis, and an efficient amplification primer of the tridacnidae shellfish mitochondria COI gene sequence is sieved. The efficient amplification primer is characterized in that CQF is 5'-GATATTGGAACCCTTTACTT-3'; and CQR is 5'-TTCAGGATTGTCCCAAGAATCA-3'. By using the tridacnidae shellfish mitochondria COI primer, the target fragments of different specifies of tridacnidae shellfishes can be efficiently amplified; and the tridacnidae shellfish mitochondria COI primer has important significance for identifying tridacnidae species and carrying out phylogenetic analysis by using a molecular method, and can provide reliable technical guidance for developing and protecting tridacnidae germplasm resources by using the molecular means.

Description

technical field [0001] The invention relates to a primer for amplifying the mitochondrial COI [the cytochrome c oxidase I] gene of Tridacnaceae shellfish. Background technique [0002] Tridacna family (Corbiculacea) belongs to Bivalvia, Heterodonta, and Veneroida. It is the largest bivalve mollusk in the world, and it is mostly distributed in the Indian Ocean and the Pacific Ocean. There are many coral reefs near the low tide areas in tropical seas such as Indonesia, Malaysia and Australia, and they are also distributed in Hainan Province and the South China Sea Islands in my country. There are 2 genera and 6 species in the world, among which giant clams, giant clams and giant clams are listed as rare and protected animals in the world. Tridacna species have been favored by people since ancient times, ranking first among the "Seven Treasures of Buddhism". Moreover, giant clam meat is delicious and nutritious. It is a valuable seafood and has certain medicinal value and orna...

Claims

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Application Information

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