General purpose primer for cloning bacteria gyrase-gyrB gene

A general primer, gyrase technology, applied in sugar derivatives, organic chemistry, DNA/RNA fragments, etc., can solve the problem of limited amount of genetic information, and achieve the effect of strong specificity and sensitivity

Inactive Publication Date: 2007-08-08
CENT SOUTH UNIV
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  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

[0008] The purpose of the present invention is to provide a new universal primer of gyrB gene, which solves the problem that the amount of genetic information contained in existing gyrB gene primers is limited

Method used

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  • General purpose primer for cloning bacteria gyrase-gyrB gene
  • General purpose primer for cloning bacteria gyrase-gyrB gene

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Embodiment 2

[0040]Through these two primers, acidophilic Thiobacillus ferrooxidans (ATCC 23270 and DQ062115 (the 16S rDNA sequence accession number of the strain is DQ672266)) of the genus Acidithiobacillus (Acidithiobacillus), Leptospirillum (Leptospirillum ) in five strains of Leptospira ferrooxidans (16S rDNA sequence accession numbers of these strains are: DQ343299, EF025341, EF025340, EF025337, DQ451017 and EF025342) and Leptospira ferrousoxidans (ATCC 53992), sphingosine Sphingomonas XJ-1 of the genus Sphingomonas (the 16S rDNA sequence accession number of the strain is DQ672266) and a strain of the genus Acidiphilum (ATCC 15859), a total of 4 genera and 11 strains were amplified from the genomic DNA gyrB gene (as shown in Table 2).

[0041] Table 2 utilizes the gyrB gene of the bacterial strain of primer amplification of the present invention

[0042] Genus and strain name

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Abstract

The invention discloses a general primer of cloned bacterium spiral-accelerating enzyme-gyrB gene, whose nucleic sequence of upstream primer is 5'-CC(ATGC)GG(ATGC)ATG TA(CT) AT(ATC)GG-3' and the nucleic sequence of downstream primer is 5'-CAT (CT) TC (ATGC) CC (ATGC) A (AG) (ATGC) CC (CT) TT (AG) (AT) A (ATGC) C (GT) (CT) TG-3'), wherein the whole sequence of PCR product approximates to the whole sequence of gyrB gene, which possesses stronger specificity and sensitivity.

Description

technical field [0001] The invention relates to the field of microbial phylogenetic molecular markers, in particular to a universal primer for bacterial gyrase-gyrB gene. Background technique [0002] In modern microbial taxonomy, 16S rDNA has been widely used to study the phylogenetic status of bacteria. not enough. The gyrB gene is the gene encoding the B subunit of DNA gyrase (a prokaryotic type II topoisomerase). As a new molecular marker, the gene has been widely used in the identification of bacteria and fungi at the species level and natural Detection of microorganisms under environmental conditions. Studies on the taxonomic identification of Aeromonas, Pseudomonas and Shewanella have proved that the molecular taxonomic index of the gyrB gene has a certain Strong feasibility. The reason, on the one hand, is that this gene has a higher base substitution frequency than 16S rDNA. The average base substitution rate of 16S rDNA is 1% every 50 million years, while gyrB ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07H21/04C12N15/52
Inventor 邱冠周夏金兰张成桂高健丁建南
Owner CENT SOUTH UNIV
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