Scarab Genomics, LLC

Wisconsin Alumni Research Foundation

Richter Gedeon Vegyeszeti Gyar Rt

Gyorgy Posfai

Frederick R Blattner

Guy Plunkett

David A Frisch

John Walter Campbell

Christopher D Herring

Jeremy D Glasner

Robert E Novy

Eric A Steffen

Hyunsic Choi

Balint Csorgo

Terrance M Henker

Charles Landry

Pal Venetianer

Imre Boros

Christopher R Blattner

Jose A Sanchez

Trevor Twose

Cristopher R Blattner

Charles F Landry

Guy D Plunkett

Twose Trevor

Trevor Martin Twose

Rediced genome or native K12 strainbacteria comprising expression vectors encoding a recombinant CRM197 protein and their use in the production of CRM 197 is provided. The CRM 197 protein may be fused to a signal sequence that directs the expressed CRM197 protein to the periplasm of thehost.

Enhanced production of recombinant CRM197 in

Reduced genome bacteria with improved genetic stability are provided. Also provided are methods of producing polypeptides using the reduced genome bacteria with improved genetic stability.

Reduced genome bacteria with improved genetic stability

bacteria comprising genetic modifications to enhance fermentability and production of protein and nucleic acids are provided.

Bacteria with improved metabolic capacity

Reduced genome strains ofMGl 655 are described. In various embodiments, the strains have one or more of equal or improved growth rate, transformation efficiency, protein expression, DNA production, DNA yield and/or DNA quality compared to the parental strain and commercially available strains.

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The present invention provides a bacterium having a genome that is genetically engineered to be smaller than the genome of its native parent strain. A bacterium with a smaller genome can produce a commercial product more efficiently. The present invention also provides methods for deleting genes and other DNA sequences from a bacterial genome. The methods provide precise deletions and seldom introduces mutations to the genomic DNA sequences around the deletion sites. Thus, the methods can be used to generate a series of deletions in a bacterium without increasing the possibility of undesired homologous recombination within the genome. In addition, some of the methods provided by the present invention can also be used for replacing a region of a bacterial genome with a desired DNA sequence.

Prophage element-free bacteria

Reduced genome strains ofMG1655 are described. In various embodiments, the strains have one or more of equal or improved growth rate, transformation efficiency, protein expression, DNA production, DNA yield and/or DNA quality compared to the parental strain and commercially available strains.

A bacteria lacking genomic and non-genomic IS elements is provided. The bacteria may be more stable and useful for the production of amino acids, polypeptides, nucleic acids and other products.

Insertion sequence-free bacteria

The present invention provides a bacterium having a genome that is genetically engineered to be at least 2 to 14% smaller than the genome of its native parent strain. A bacterium with a smaller genome can produce a commercial product more efficiently. The present invention also provides methods for deleting genes and other DNA sequences from a bacterial genome. The methods provide precise deletions and seldom introduces mutations to the genomic DNA sequences around the deletion sites. Thus, the methods can be used to generate a series of deletions in a bacterium without increasing the possibility of undesired homologous recombination within the genome. In addition, some of the methods provided by the present invention can also be used for replacing a region of a bacterial genome with a desired DNA sequence.

Bacteria with reduced genome

The present invention discloses that a bacterium having a genome that is genetically engineered to be at least 10% smaller than the genome of its native parent strain has better transformation competence. Specificstrains, having significantly reduced genome sizes, are disclosed which are highly transformation competent. A medium and methodology is taught which enables transformation efficiencies to be increased further.

Competent bacteria

Plasmid vectors and a process for the preparation thereof are disclosed having the same replication region as pBR322, but which have a substantially higher copy number per cell. The plasmid vectors contain a G.fwdarw.T point mutation at base pair 3074 of the pBR322 sequence which holds back synthesis of the repressor of plasmid replication, and also contain either no tetracycline resistance gene or the tetracycline resistance gene in an inactivated or weakened state.

Growing community of inventors

Szeged, Hungary

Gyorgy Posfai