Optimized genetic tools for modifying bacteria

A genetic tool, bacterial technology, applied in the direction of bacteria, the use of vectors to introduce foreign genetic material, microbial-based methods, etc., can solve problems such as toxicity limitations

Pending Publication Date: 2022-04-05
INST FR DU PETROLE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Toxicity of this system is limited by placing the cas9 and / or gRNA expression cassette under the control of an inducible promoter

Method used

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  • Optimized genetic tools for modifying bacteria
  • Optimized genetic tools for modifying bacteria
  • Optimized genetic tools for modifying bacteria

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0034] Despite more than a century of use in industry, knowledge of solventogenic bacteria, especially those belonging to the genus Clostridium, is still limited by the difficulties encountered in their genetic modification. For example, naturally isopropanol-producing Clostridium bacteria, which typically have in their genome an adh gene encoding a primary / tertiary alcohol dehydrogenase that reduces acetone to isopropanol, are genetically and functionally distinct from those capable of producing Bacteria for ABE fermentation.

[0035] Advantageously, in the context of the present invention, the inventors have successfully transformed and genetically modified a bacterium of the genus Clostridium that naturally produces isopropanol, the bacterium Clostridium bejejejerinckii DSM 6423, and the reference strain Clostridium acetobutylicum DSM 792.

[0036] Some of the work described in the experimental section was performed within a strain capable of IBE fermentation, Clostridium ...

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Abstract

The present invention relates to the transformation and genetic modification of bacteria belonging to the Phylum Thallungiae. Thus, the invention relates to methods, tools and kits that allow such genetic modifications to proceed, in particular to nucleic acid sequences for promoting the transformation of said bacteria, comprising i) all or part of the sequence SEQ ID NO: 126, and ii) a sequence that allows modification of the genetic material of the bacterium and / or allows partial or complete absence of DNA sequences in genetic material present in a wild-type version of the bacterium to be expressed within the bacterium. The invention also relates to the genetically modified bacteria obtained and to the use thereof, in particular for the production of solvents, preferably on an industrial scale.

Description

[0001] The present invention relates to bacteria, in particular bacteria belonging to the phylum Firmicutes, generally solventogenic bacteria such as bacteria of the genus Clostridium, preferably having a bacterial chromosome in the wild state and at least one DNA molecule different from the chromosomal DNA (or Bacterial transformation and genetic modification of both natural plasmids). The present invention therefore relates to methods, tools and kits allowing such genetic modifications, in particular to nucleic acid sequences for facilitating the transformation of said bacteria, said sequences comprising i) all or part of the sequence SEQ ID NO: 126 , and ii) a sequence that allows the modification of the genetic material of the bacterium and / or the expression in the bacterium of a DNA sequence that is partially or completely absent in the genetic material present in the wild-type version of the bacterium. The invention also relates to the genetically modified bacteria obtaine...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/21C12N15/74C12N15/113C12N9/22C12N9/10C12N15/10C12R1/145
CPCC12N15/102C12N15/74C12N2310/20C12R2001/145C12N1/205Y02E50/10C12N9/22C12N15/113C12N15/111C12N15/902
Inventor 尼古拉斯·洛普斯费雷拉雷米·霍克弗朗索瓦·瓦塞尔斯
Owner INST FR DU PETROLE
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