Dual-promoter inducible secretable shuttle plasmid and construction method thereof

A dual-promoter, shuttle plasmid technology, applied in the field of bioengineering, can solve problems such as high cost, inability to large-scale production, and small amount of bactericidal peptides
CN101948865AInactive Publication Date: 2011-01-19FUDAN UNIV
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
FUDAN UNIV
Publication Date
2011-01-19
Estimated Expiration
Not applicable · inactive patent

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Abstract

The invention belongs to the field of biological genetic engineering, and provides a dual-promoter inducible secretable shuttle plasmid. The shuttle plasmid consists of an enzyme cutting Escherichia coli pET-28a plasmid and an enzyme cutting bacillus subtilis pE194 plasmid, wherein a lysozyme-antibacterial peptide functional fragment and an antibacterial peptide gene are inserted into the enzyme cutting Escherichia coli pET-28a plasmid. The invention also provides a method for constructing the dual-promoter inducible secretable shuttle plasmid. The shuttle plasmid is transferred to a prokaryote to express proteins such as toxalbumin, cecropin and the like in a fusion mode without toxicity to a host. The shuttle plasmid constructed by the method can be induced by IPTG or lactose to improve gene expression level; the shuttle plasmid is also suitable for exocrine expression, and a target protein for the exocrine expression is convenient to process and is not polluted by endotoxin; and because the shuttle plasmid has dual functional promoters, different genes can be inserted in appropriate correct directions for gene expression.
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Description

Technical field:

[0001] The present invention relates to the field of bioengineering, in particular to a shuttle plasmid, in particular to a dual-promoter inducible and secretable shuttle plasmid and a construction method thereof. Background technique:

[0002] In the field of bioengineering, in order to effectively express cloned genes, it is often necessary to construct expression vectors. Escherichia coli expression system is the most commonly used one in genetic engineering research. Although it cannot perform post-translational processing like the eukaryotic system, the genetic background of Escherichia coli is relatively clear, easy to operate, short in growth cycle, and economically safe, so it is still the most widely used and irreplaceable system. It has become an indispensable and important tool in research and practical application. Some important exogenous genes have been highly expressed in Escherichia coli, accounting for 30% or even 40% of the total bacteria...

Claims

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