Construction of bacillus subtilis integrated vector capable of being cloned by using homologous recombination and application thereof

A Bacillus subtilis, homologous recombination technology, applied in the biological field, can solve the problems of not being widely used, inconvenient cloning of foreign genes, and difficult formation of inclusion bodies, etc.
CN101948866AInactive Publication Date: 2011-01-19南宁邦尔克生物技术有限责任公司
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Patent Information

Authority / Receiving Office
CN · China
Current Assignee / Owner
南宁邦尔克生物技术有限责任公司
Publication Date
2011-01-19
Estimated Expiration
Not applicable · inactive patent
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Abstract

The invention discloses construction of a bacillus subtilis integrated vector capable of being cloned by using homologous recombination and application thereof. The construction of the bacillus subtilis integrated vector capable of being cloned by using homologous recombination and the application thereof belong to the technical field of biology. The vector pMLK-BN can transfer an exogenous gene or a promoter cloned on a common cloning vector puc18, puc19 or puc57 to self by a homologous recombination method so as to integrally express exogenous protein in bacillus subtilis or test the promoting effect of the promoter. Compared with a common connecting cloning method, the cloning method of homologous recombination is simple and efficient; and the pMLK-BN can be effectively used as a choice for cloning the exogenous gene or the promoter to the bacillus subtilis to perform integrated expression.
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Description

technical field

[0001] The invention belongs to the field of biotechnology, in particular to the construction and application of a vector capable of cloning by homologous recombination and integrating and expressing exogenous DNA fragments in Bacillus subtilis. Background technique

[0002] Bacillus subtilis is non-pathogenic, does not produce toxins and thermosensitive proteins, is a food-safe strain, and is classified as a food-grade microorganism. Moreover, as an expression system, Bacillus subtilis has the following advantages: 1. It has a strong protein secretion function. It does not need to break cells to extract proteins. It only needs to simply process the fermentation supernatant to obtain a relatively pure target protein. A variety of exogenous proteins have been secreted and expressed in Bacillus subtilis. 2 There is no obvious codon preference, and the expression product is not easy to form inclusion bodies. 3 The fermentation conditions are simple. Therefore...

Claims

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