Bacillus licheniformis for multiplying operon bacABC copy number and knocking off recA gene and establishment method of bacillus licheniformis

A Bacillus licheniformis, copy number multiplication technology, applied in the direction of microorganism-based methods, chemical instruments and methods, biochemical equipment and methods, etc., can solve the difficulty of increasing the copy number of the operon bacABC, the inability to construct plasmid expression vectors, and strain maintenance Unfavorable copy number stability and other issues, to achieve the effect of increasing potency

Active Publication Date: 2015-01-21
LIFECOME BIOCHEM
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Problems solved by technology

However, due to the operon bacABC The gene sequence is too long, about 42kb, so it is impossible to construct a plasmid expression vector containing such a large gene fragment, and it is impossible to realize the operon by constructing an episomal or integrated plasmid expression vector bacABC Copy number doubling, therefore, the operon bacABC C

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  • Bacillus licheniformis for multiplying operon bacABC copy number and knocking off recA gene and establishment method of bacillus licheniformis
  • Bacillus licheniformis for multiplying operon bacABC copy number and knocking off recA gene and establishment method of bacillus licheniformis
  • Bacillus licheniformis for multiplying operon bacABC copy number and knocking off recA gene and establishment method of bacillus licheniformis

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Embodiment Construction

[0039] an operon bacABC Copy number doubling and knockout recA Bacillus licheniformis gene, said bacterial strain is Bacillus licheniformis DW2△ recA / bacABCs (Referred to as: Bacillus licheniformis DW2△ recA / bacABCs ), the strain is preserved in the China Center for Type Culture Collection in Wuhan, with the preservation number CCTCC NO: M2014251, and the preservation date is June 12, 2014. The genome of this strain also has an operon bacABC Copy number doubling and knockout recA Gene dual characteristics, strong ability to produce bacitracin.

[0040] an operon bacABC Copy number doubling and knockout recA The construction method of the bacillus licheniformis strain of gene, comprises the following steps:

[0041] (1) Plasmid T2(2)-ori (the schematic diagram of its structure, such as figure 1 shown) after restriction DNA endonuclease Xba I and Bam H I The linear T2(2)-ori plasmid was obtained by double enzyme digestion, and the linear T2(2)-ori p...

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Abstract

The invention provides bacillus licheniformis DW2delta recA bacABCs for multiplying the operon bacABC copy number and knocking off a recA gene. The strain is preserved in the China Center for Type Culture Collection in wuhan on June 12th, 2014, and the preservation number is CCTCC NO:M2014251. The genome of the strain also has double characteristics of multiplying the operon bacABC copy number and knocking off the recA gene, and moreover when the bacillus licheniformis DW2delta recA bacABCs for multiplying the operon bacABC copy number and knocking off the recA gene is used in bacitracin production, the yield of bacitracin can be increased by 11.5%.

Description

technical field [0001] The present invention relates to an operon bacABC Copy number doubling and knockout recA Genetically engineered strains of Bacillus licheniformis. Background technique [0002] Bacitracin is a light yellow or off-white powder, odorless and bitter in taste. It is a polypeptide antibiotic composed of 10 amino acids and 12 amino acid residues, which can effectively inhibit Gram-positive pathogenic bacteria and some Gram-negative bacteria. Bacitracin has the advantages of almost no absorption in the intestine, rapid excretion, and no residue in the body, so it is widely added to feed. [0003] At present, the main production strain of bacitracin is Bacillus licheniformis ( Bacillus licheniformis ) and Bacillus subtilis ( Bacillus subtilis ), which is produced by microbial fermentation. [0004] Bacitracin is formed by condensation of amino acids through a sulfur template mechanism, while the operon bacABC It is the gene encoding three key enzymes...

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Application Information

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IPC IPC(8): C12N1/21C12N15/75C12P21/00C12R1/10
CPCC07K7/58C12N9/00C12N15/52
Inventor 陈守文彭炳王冬李阳祁高富孔素芬陈惠超陈晓斌
Owner LIFECOME BIOCHEM
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