Novel plasmid vector and transformant capable of carrying plasmid stably

Abstract

The object is to develop: a novel vector, preferably a novel vector which can be carried in a cell of a bacterium belonging to the genus Ralstonia, Cupriavidus or Wautersia stably in the absence of selection pressure for an antibiotic and has no conjugal transfer property; a strain capable of stably producing a polyhydroxyalkanoate using the vector; and a method for production of a polyhydroxyalkanoate using the strain. Disclosed is a novel recombinant vector having a DNA replication initiation segment which can function in a cell of a bacterium belonging to the genus Ralstonia, Cupriavidus or Wautersia, particularly a recombinant vector having both a DNA replication initiation segment which can function in a cell of a bacterium belonging to the genus Ralstonia, Cupriavidus or Wautersia and a segment which can stabilize the recombinant vector (a par segment). By using a transformant produced using the vector, the vector can be carried in a bacterium stably and a polyhydroxyalkanoate can be produced with good efficiency.

Description

technical field The present invention relates to new vectors. In addition, the present invention belongs to the technical field of plasmid stabilization. More specifically, the present invention relates to the ability to stably exist in the genus Ralstia, Copperphyllum, or Waterella, which are hydrogen bacteria and well-known PHB-synthesizing bacteria. The recombinant vector, as well as the strain transformed by the vector and the commercial production of polyhydroxyalkanoate using the strain. Background technique When recombinant DNA technology is actually used to produce a target substance by microorganisms, there is generally a problem of instability of the recombinant plasmid. Cloning and expression vectors commonly used in laboratories are usually multi-copy plasmids, and their stable inheritance to offspring is ensured by introducing multiple plasmids into each cell genome (refer to Non-Patent Document 1). However, if a plasmid is used to introduce a foreign gene, t...

AI Technical Summary

Problems solved by technology

If culture is performed by applying selective pressure with antibiotics, there are problems as follows: (1) the use of antibiotic-resistant strains causes harm to the environment, (2) the amount of antibiotics necessary for culture will significantly increase production costs, (3) antibiotics Production of substances unsuitable for use in the treatment of humans and animals

However, the aforementioned stabilization of plasmids using the par region is not applicable to bacteria belonging to the genus Ralstia, Copperphyllum, and Wateria, and it is unclear whether conventional system construction methods and constructed systems are effective for plasmid stabilization. Does stabilization actually work?

Images