Bacillus megaterium co-expression vector for nitrite reductase and construction method thereof

A technology of Bacillus megaterium and co-expression vector, applied in the field of genetic engineering, can solve the problems of failing to meet the requirements of repairing the environment and low expression of nitrite reductase
CN103614404AInactive Publication Date: 2014-03-05SHANGHAI JIAO TONG UNIV
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Patent Information

Authority / Receiving Office
CN · China
Current Assignee / Owner
SHANGHAI JIAO TONG UNIV
Publication Date
2014-03-05
Estimated Expiration
Not applicable · inactive patent

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Abstract

The invention relates to a bacillus megaterium co-expression vector for nitrite reductase and a construction method thereof, belonging to the technical field of genetic engineering. According to the co-expression vector, all genome sequences of bacillus megaterium are determined and serve as a template, gene sequences containing restriction enzyme cutting sites, namely a large subunit (Bm-Nas D) and a small subunit (Bm-Nas E) of nitrite reductase, are amplified according to designed PCR (Polymerase Chain Reaction) primers and are sequentially connected to the co-expression vector pETDuet-1, and positive clones of Escherichia coli DH5alpha containing target gene fragments are selected by a colony PCR technology. The co-expression vector has the advantages that the defect in the prior art that the use effect is seriously restricted due to the fact that the expression level of the gene sequence of the small subunit of nitrite reductase in natural bacillus megaterium is very low is overcome, and the genetic expression level is increased greatly.
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Description

technical field

[0001] The present invention relates to a method in the technical field of genetic engineering, specifically a nitrite reductase small subunit gene sequence, a recombinant vector containing the gene, a recombinant bacterium, and the expression product of the gene produced by the recombinant bacterium and its application. Background technique

[0002] From an environmental point of view, the accumulation of N elements can cause waste of fertilizers and affect crop growth and soil utilization. At the same time, it can cause agricultural non-point source pollution through farmland surface runoff and farmland leakage; from the perspective of food safety, the human body Nitrate (NO 3 –) easily reduced to nitrite (NO 2 –), nitrite can oxidize the oxygen-carrying low iron hemoglobin in the cells to metherin without oxygen-carrying capacity and cause tissue hypoxia. Not only that, the oxidized methemoglobin will also reduce the oxygenation of the oxygen-carrying T...

Claims

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