Virulence attenuated bacteria based protein delivery

A technology of bacteria and proteins, applied in the direction of bacteria, bacterial peptides, peptide/protein components, etc.

Pending Publication Date: 2019-10-11
UNIVERSITY OF BASEL
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The development of bacterial strains capable, for example, of delivering intrabacterially produced cargo to its site of action (i.e. outside the bacterium) within cells (such as cancer cells) remains a major challenge.

Method used

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  • Virulence attenuated bacteria based protein delivery
  • Virulence attenuated bacteria based protein delivery
  • Virulence attenuated bacteria based protein delivery

Examples

Experimental program
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Embodiment 1

[0283] A) Materials and methods

[0284] Bacterial strains and growth conditions. The strains used in this study are listed in Figure 3A to M. E. coli Top10 for plasmid purification and cloning, and E. coli Sm10λpir for conjugation, and E. coli BW19610 for propagation of pKNG101 31 Routine growth was performed on LB agar plates and in LB media at 37°C. Expression vectors were selected using ampicillin at a concentration of 200 μg / ml (Yersinia) or 100 μg / ml (E. coli). Streptomycin was used at a concentration of 100 μg / ml to select for suicide vectors. Yersinia enterocolitica MRS40 (O:9, biotype 2) 20 (a non-ampicillin-resistant E40-derivative) 19 and its derivative strains were routinely grown on brain heart infusion (BHI; Difco) at room temperature. Nalidixic acid (35 μg / ml) was added to all Y. enterocolitica strains and all Y. enterocolitica asd strains were additionally supplemented with 100 μg / ml meso-2,6-diamino Pimelic acid (mDAP, Sigma Aldrich). Salmonella ente...

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Abstract

The present invention relates to recombinant virulence attenuated Gram-negative bacterial strains and its use in a method of treating cancer in a subject.

Description

technical field [0001] The present invention relates to recombinant attenuated Gram-negative bacterial strains and their use in methods of treating cancer in a subject. Background technique [0002] Bacteria have evolved different mechanisms to inject proteins directly into target cells 1 . bacteria such as Yersinia, Shigella and Salmonella 2 The type III secretion system (T3SS) used by bacteria functions like a nano-injector for injecting so-called bacterial effector proteins into host cells. [0003] T3S has been used to deliver hybrid peptides and proteins into target cells. Heterologous bacterial T3SS effectors have been delivered in case the bacteria studied are difficult to obtain genetically (e.g. Chlamydia trachomatis). Reporter proteins are often fused to possible T3SS secretion signals to study the requirement for T3SS-dependent protein delivery, such as Bordetella pertussis adenylate cyclase, murine DHFR, or a phosphorylatable tag. Peptide delivery is mainly ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/195C12N15/70C12N15/74C07K14/52
CPCC07K14/24C07K2319/02C07K2319/036C07K2319/21C07K2319/41C07K2319/50C12N15/70C12N15/74A61P35/00Y02A50/30A61K38/162A61K38/164A61K2039/522A61K38/00C12N1/20C12N1/36
Inventor 西蒙·伊泰格马利斯·阿姆斯特茨克里斯托夫·卡斯帕
Owner UNIVERSITY OF BASEL
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