Mitochondrion complete genome sequence of hucho taimen and amplification primers thereof

A whole genome, sequence amplification technology, applied in genetic engineering, plant genetic improvement, recombinant DNA technology, etc., can solve the problems of slowness, high temperature resistance, impact on cost and yield, etc.

Inactive Publication Date: 2010-06-16
HEILONGJIANG RIVER FISHERY RES INST CHINESE ACADEMY OF FISHERIES SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, due to its serious self-mutilation, intolerance to high temperature, intolerance to low oxygen, and slow growth in the low temperat

Method used

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  • Mitochondrion complete genome sequence of hucho taimen and amplification primers thereof
  • Mitochondrion complete genome sequence of hucho taimen and amplification primers thereof

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specific Embodiment approach 1

[0043] Specific embodiment 1: In this embodiment, the complete genome sequence of the salmon mitochondria is shown in SEQ ID NO: 1.

[0044] The whole genome sequence of the salmon mitochondria is 16797bp in total, including 22 tRNA genes, 1 12S rRNA gene, 1 16S rRNA gene and 13 protein-coding genes. The corresponding sequence positions of the genes and other structures in the salmon mitochondria are shown in Table 1 Show.

[0045] Table 1

[0046] gene / component

[0047] gene / component

[0048] gene / component

specific Embodiment approach 2

[0049] Specific embodiment two: In this embodiment, there are 33 pairs of amplification primers for the whole genome sequence of the salmon mitochondria;

[0050] The forward primer of the first pair of primers for the whole genome sequence amplification of jelly salmon is 5'-AGAGCGCCGGTCTTGTAATC-3', and the reverse primer is 5'-GCTAGCGGGACTTTCTAGGGTC-3';

[0051] The forward primer of the second pair of primers for the whole mitochondrial genome sequence amplification of jelly salmon is 5'-TGAATTCCAGAGAACCCATGT-3', and the reverse primer is 5'-TAAAACAAGGGGTGGTGAGG-3';

[0052] The forward primer of the third pair of primers for the whole genome sequence amplification of jelly salmon is 5'-CAAAGGCTTGGTCCTGACTT-3', and the reverse primer is 5'-TAAAACAAGGGGTGGTGAGG-3';

[0053] The forward primer of the fourth pair of primers for the whole genome sequence amplification of jelly salmon is 5'-TGATATCCGCCAGGGAACTA-3', and the reverse primer is 5'-CCCTTGCGGTACTTTTTCTG-3';

[0054] ...

specific Embodiment approach 3

[0084] Specific embodiment three: In this embodiment, 33 pairs of primers in specific embodiment two are used to amplify and obtain the whole genome sequence of jellyfish mitochondria according to the following steps:

[0085] 1. Extraction of mitochondrial genomic DNA from salmon

[0086] A. Take 200 mg fresh or -20°C preserved salmon muscle and liver, wash and grind with sterile water, and then extract mitochondria (see the operation manual of commercial mitochondrial extraction kit for the operation steps and parameters of mitochondrial extraction); B, Add SDS with a mass concentration of 20% to the centrifuge tube containing the mitochondrial suspension, so that the final concentration of SDS reaches 0.8%, and then bathe in water at 37°C for 15 minutes; C. Add phenol chloroform equal to the volume of the liquid in the centrifuge tube Extract, shake gently until milky white, and stand on ice for 10 minutes; D. Centrifuge at 10,000g for 10 minutes at 4°C, and transfer the su...

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Abstract

The invention discloses a mitochondrion complete genome sequence of hucho taimen and amplification primers of the mitochondrion complete genome sequence, which relate to the mitochondrion complete genome sequence and the amplification primer of the mitochondrion complete genome sequence. The mitochondrion complete genome sequence of the hucho taimen determined by the invention provides a material basis for conservation genetics, evolutionary genetics and molecular marker assisted selection of good breeding species of the hucho taimen. The mitochondrion complete genome sequence of the hucho taimen is as shown by SEQ ID NO:1. The mitochondrion complete genome sequence of the hucho taimen comprises 33 pairs of the amplification primers. The mitochondrion complete genome sequence of the hucho taimen and the amplification primers of the mitochondrion complete genome sequence provide the material basis for the conservation genetics, the evolutionary genetics and the molecular marker assisted selection of the good breeding species of the hucho taimen.

Description

technical field [0001] The invention relates to a whole mitochondrial genome sequence and amplifying primers thereof. Background technique [0002] Mitochondria is an important organelle in the cell. It has its own set of genomes. Due to the characteristics of maternal inheritance in fish, the mitochondrial genome lacks a repair mechanism during the replication process, resulting in the cumulative variation rate of the sequence being much greater than that of the nuclear genome. And other characteristics, making it a very effective tool for conservation biology and evolutionary biology research. At the same time, because mitochondria play an important role in energy production and energy transfer in cells, base mutations or deletions in their genomes can cause major mitochondrial diseases (such as human mitochondrial muscle disease and mitochondrial brain muscle disease, etc.), and their genomes are multiple Morphology is related to the economic traits of species, so the st...

Claims

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Application Information

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IPC IPC(8): C12N15/12C12N15/11C12Q1/68
Inventor 佟广香匡友谊尹家胜孙效文
Owner HEILONGJIANG RIVER FISHERY RES INST CHINESE ACADEMY OF FISHERIES SCI
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