Shuttle plasmid carrier, and construction method and application thereof

A technology of shuttle plasmid and vector, applied in the field of shuttle plasmid vector and its construction, can solve problems such as difficulties in foreign genes
CN107760705AActive Publication Date: 2018-03-06ZHEJIANG UNIV OF TECH
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
ZHEJIANG UNIV OF TECH
Publication Date
2018-03-06

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Abstract

The invention discloses a shuttle plasmid carrier, and a construction method and application thereof. The shuttle plasmid carrier is formed by linearizing bacillus subtilis plasmid pWB980 and inserting into a carrier pMD19-T Simple Vector. The shuttle carrier plasmid can be copied in escherichia coli and paenibacillus polymyxa. In the paenibacillus polymyxa, the genetic operation system is not yetmature, the conversion steps are tedious and the conversion efficiency is relatively low; and in the escherichia coli, gene cloning operation is very convenient. The plasmid carrier can complete exogenous gene cloning and recombinant plasmid construction in the escherichia coli, related gene function research and genetic breeding work are developed in the paenibacillus polymyxa conveniently, andthe conversion rate can reach 7.83 + / -1.54*10<5>CFU / ug DNA.
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Description

(1) Technical field

[0001] The invention relates to a shuttle plasmid vector and its construction method and application. (2) Background technology

[0002] Paenibacillus polymyxa is a rod-shaped, spore-forming, Gram-positive bacterium that is mostly isolated from plant rhizosphere soil. In recent years, due to its remarkable plant growth-promoting effect and antibacterial effect, it is generally regarded as a kind of plant rhizosphere growth-promoting bacteria. Paenibacillus polymyxa can produce a variety of antagonistic substances that inhibit the growth of bacteria and fungi, such as polymyxin, cyclobacterin, cholipidin, polypeptide mycetin, oryvalerin and fusaricidin, etc. antibiotics, and because some antibiotics are synthesized by non-ribosomal peptide synthetases, this has attracted the attention of more and more researchers.

[0003] Plasmid pWB980 is a Bacillus subtilis expression vector constructed by Wu et al. in 1999 based on the transformation of pUB110. This ...

Claims

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