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Literature summary extracted from

  • Li, J.; Sun, J.; Gao, X.; Wu, Z.; Shang, G.
    Coupling ssDNA recombineering with CRISPR-Cas9 for Escherichia coli DnaG mutations (2019), Appl. Microbiol. Biotechnol., 103, 3559-3570 .
    View publication on PubMed

Protein Variants

EC Number Protein Variants Comment Organism
2.7.7.101 K580A site-directed mutagenesis, the mutant shows 1.6fold recombineering efficiency compared to wild-type Escherichia coli
2.7.7.101 additional information ssDNA recombineering and CRISPR-Cas9 are combined for the generation of DnaG variants. The tightly regulated Red operon expression cassette and tightly regulated Cas9 expression cassette are integrated into one chloroamphenicol resistance, p15A replicon-based vector. A self-curing, kanamycin resistance, p15A replicon-based plasmid is applied for the plasmid elimination after genome editing. The genome editing efficiency is as high as 100%. The recombineering efficiency of the strains harboring the DnaG variants is assayed via the kanamycin resistance gene repair as well as the chromosomal gene deletion experiments. Genotype analysis. THe Q576A/K580A double variant cannot be generated, Q576A/K580A is lethal to Escherichia coli Escherichia coli
2.7.7.101 Q576A site-directed mutagenesis, the mutant shows 2.8fold recombineering efficiency compared to wild-type Escherichia coli
2.7.7.101 Q576T/K580A site-directed mutagenesis, the mutant shows 3.6-7.0fold recombineering efficiency compared to wild-type Escherichia coli
2.7.7.101 Q576V/K580A site-directed mutagenesis, the mutant shows 5.4-7.5fold recombineering efficiency compared to wild-type Escherichia coli

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.7.7.101 ssDNA + n NTP Escherichia coli
-
 ssDNA/pppN(pN)n-1 hybrid + (n-1) diphosphate
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.7.7.101 Escherichia coli P0ABS5
-
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.7.7.101 additional information enzyme recombineering efficiency assay via gene deletion Escherichia coli ?
-
-
2.7.7.101 ssDNA + n NTP
-
 Escherichia coli ssDNA/pppN(pN)n-1 hybrid + (n-1) diphosphate
-
 ?

Synonyms

EC Number Synonyms Comment Organism
2.7.7.101 DnaG
-
 Escherichia coli
2.7.7.101 DnaG primase
-
 Escherichia coli