Complete sequence cloning and evolutionary analysis on boar mitochondrial genome in Heilongjiang Province

A mitochondrial genome, Heilongjiang Province technology, applied in genetic engineering, plant genetic improvement, recombinant DNA technology, etc., can solve the problem of no mitochondrial genome sequence records of wild boars in Heilongjiang Province.

Inactive Publication Date: 2010-06-09
刘娣 +2
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  • Abstract
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Problems solved by technology

At present, on the website of the National Center for Biotechnology Information (NCBI) in the United States, the complete mitochondrial genome sequences of more than 50 wild boar species around the world can be found, but there is no record of the mitochondrial genome sequence of wild boars in Heilongjiang Province
This has certain limitations for the study of the origin of domestic pigs in Heilongjiang Province and even my country

Method used

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  • Complete sequence cloning and evolutionary analysis on boar mitochondrial genome in Heilongjiang Province
  • Complete sequence cloning and evolutionary analysis on boar mitochondrial genome in Heilongjiang Province
  • Complete sequence cloning and evolutionary analysis on boar mitochondrial genome in Heilongjiang Province

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Experimental program
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Embodiment Construction

[0011] 1. Experimental materials

[0012] 1. Experimental animals

[0013] One wild boar from Heilongjiang Province used in this experiment was captured in the Xiaoxing’an Mountains. Part of the liver tissue was collected after slaughter and stored in liquid nitrogen.

[0014] 2. Strains

[0015] Escherichia coli (Escherichia coli) DH5α was purchased from Beijing Dingguo Biotechnology Co., Ltd.

[0016] 3. Main related reagents for molecular cloning

[0017] (1) BamHI, EcoRI, HindIII and other restriction enzymes, rTaq enzyme, dNTP, DL2000Marker, and pMD-18T vector ligation kits were all purchased from Dalian Bao Biological Engineering Co., Ltd.

[0018] (2) Gel recovery (small amount) kit and plasmid (small amount) extraction kit were purchased from Shanghai Huashun Biological Engineering Co., Ltd.

[0019] (3) Tryptone and yeast extract for bacterial culture were purchased from Oxid Company.

[0020] (4) Agarose, ethidium bromide, Tris, etc. were purchased from Sigma. ...

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Abstract

The invention designs a set of commonly-used primer combination for mitochondrial whole genome amplification of boars, adopts a multi-primer overlap method to measure the mitochondrial whole genome sequence for boars in Heilongjiang Province, performs positioning analysis on two rRNAs, 22 tRNAs, 13 protein coding genes and 1 D-loop area, and constructs a molecular phylogenetic tree for current suidae animals by using the sequence and the mitochondrial genome sequence submitted by NCBI, wherein, the two rRNAs, 22 tRNAs and 13 protein coding genes are contained in the mitochondrial whole genome sequence.

Description

technical field [0001] The invention relates to the cloning of the pig genome sequence, specifically, the complete mitochondrial genome sequence of the wild boar in Heilongjiang Province is cloned, and the molecular evolution tree of the pig family is constructed according to the existing mitochondrial genome sequence of the pig family. Background technique [0002] Mitochondria is an organelle of eukaryotic cells, has its own genome, encoding some of the proteins of the organelle. Except for the mitochondrial genome of a few lower eukaryotes, which is a linear DNA molecule (such as ciliated protozoa and green algae, etc.), it is generally a circular DNA molecule. The size of the mitochondrial genome varies widely among different species. It is known that mammals have the smallest mitochondrial genomes, fruit flies and frogs are slightly larger, yeasts are larger, and plants have the largest mitochondrial genomes. The mitochondrial gene DNA of mammals has no introns, and a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/12C12N15/10
Inventor 刘娣
Owner 刘娣
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