Erratum: Creation of a Dense Transposon Insertion Library Using Bacterial Conjugation in Enterobacterial Strains Such As Escherichia Coli or Shigella flexneri

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Erratum: Creation of a Dense Transposon Insertion Library Using Bacterial Conjugation in Enterobacterial Strains Such As Escherichia Coli or Shigella flexneri. J. Vis. Exp. (135), e6113, (2018).

Abstract

An erratum was issued for: Creation of a Dense Transposon Insertion Library Using Bacterial Conjugation in Enterobacterial Strains Such As Escherichia Coli or Shigella flexneri. The Discussion and References sections have been corrected.

The second paragraph in the Discussion section was updated from:

A significant benefit of this method is that, in theory, the user has wide latitude in the choice of enterobacterial recipient strains. This paper, as well as others11, use E. coli as a recipient strain, however the pJA1 plasmid has been used successfully with other enterobacterial recipient species such as Shigella flexneri6 and Salmonella enterica serovar Typhimurium strain SL134410. Theoretically, the γ origin of replication (oriR6Kγ) in pJA1 allows this plasmid to be maintained in a broad host range19, allowing that the recipient strain is pir+. Recently, new methods have been described that allow for construction of the pir+ in a range of enterobacterial strains20, giving additional flexibility. Additionally, the300 base pair mob region from the RP4 plasmid in pJA1 allows conjugative transfer of this plasmid to a wide range of gram negative bacterial strains19. Simply put, this method could theoretically be used with a variety of recipient strains, as long as several conditions are met: the strain is pir+, and is marked with an antibiotic resistance other than kanamycin and other than the donor strain.

to:

A significant benefit of this method is that, in theory, the user has wide latitude in the choice of enterobacterial recipient strains. This paper, as well as others11, use E. coli as a recipient strain, however the pJA1 plasmid has been used successfully with other enterobacterial recipient species such as Shigella flexneri6 and Salmonella enterica serovar Typhimurium strain SL134410. Theoretically, the γ origin of replication (oriR6Kγ) in pJA1 allows this plasmid to be maintained in a broad host range19, allowing that the recipient strain is pir-. The 300 base pair mob region from the RP4 plasmid in pJA1 allows conjugative transfer of this plasmid to a wide range of gram negative bacterial strains19. Simply put, this method could theoretically be used with a variety of recipient strains, as long as several conditions are met: the strain is pir-, and is marked with an antibiotic resistance other than kanamycin and other than the donor strain.

The References section was updated from:

  1. Blattner, F. R., Plunkett, G., et al. The Complete Genome Sequence of Escherichia coli K-12. Science. 277 (5331), 1453-1462 (1997).
  2. Alexeyev, M. F., & Shokolenko, I. N. Mini-Tn10 transposon derivatives for insertion mutagenesis and gene delivery into the chromosome of gram-negative bacteria. Gene. 160 (1), 59-62 (1995).
  3. Kvitko, B. H., Bruckbauer, S., et al. A simple method for construction of pir+ Enterobacterial hosts for maintenance of R6K replicon plasmids. BMC Res Notes. 5 (1), 157 (2012).
  4. Barrick, J. E., Yu, D. S., et al. Genome evolution and adaptation in a long-term experiment with Escherichia coli. Nature. 461 (7268), 1243-1247 (2009).
  5. Jacobs, M. A., Alwood, A., et al. Comprehensive transposon mutant library of Pseudomonas aeruginosa. PNAS. 100 (24), 14339-14344 (2003).
  6. van Opijnen, T., & Camilli, A. Transposon insertion sequencing: a new tool for systems-level analysis of microorganisms. Nature Rev Microbiol. 11 (7), 435-442 (2013).

to:

  1. Blattner, F. R., Plunkett, G., et al. The Complete Genome Sequence of Escherichia coli K-12. Science. 277 (5331), 1453-1462 (1997).
  2. Alexeyev, M. F., & Shokolenko, I. N. Mini-Tn10 transposon derivatives for insertion mutagenesis and gene delivery into the chromosome of gram-negative bacteria. Gene. 160 (1), 59-62 (1995).
  3. Barrick, J. E., Yu, D. S., et al. Genome evolution and adaptation in a long-term experiment with Escherichia coli. Nature. 461 (7268), 1243-1247 (2009).
  4. Jacobs, M. A., Alwood, A., et al. Comprehensive transposon mutant library of Pseudomonas aeruginosa. PNAS. 100 (24), 14339-14344 (2003).
  5. van Opijnen, T., & Camilli, A. Transposon insertion sequencing: a new tool for systems-level analysis of microorganisms. Nature Rev Microbiol. 11 (7), 435-442 (2013).

Protocol

An erratum was issued for: Creation of a Dense Transposon Insertion Library Using Bacterial Conjugation in Enterobacterial Strains Such As Escherichia Coli or Shigella flexneri. The Discussion and References sections have been corrected.

The second paragraph in the Discussion section was updated from:

A significant benefit of this method is that, in theory, the user has wide latitude in the choice of enterobacterial recipient strains. This paper, as well as others11, use E. coli as a recipient strain, however the pJA1 plasmid has been used successfully with other enterobacterial recipient species such as Shigella flexneri6 and Salmonella enterica serovar Typhimurium strain SL134410. Theoretically, the γ origin of replication (oriR6Kγ) in pJA1 allows this plasmid to be maintained in a broad host range19, allowing that the recipient strain is pir+. Recently, new methods have been described that allow for construction of the pir+ in a range of enterobacterial strains20, giving additional flexibility. Additionally, the300 base pair mob region from the RP4 plasmid in pJA1 allows conjugative transfer of this plasmid to a wide range of gram negative bacterial strains19. Simply put, this method could theoretically be used with a variety of recipient strains, as long as several conditions are met: the strain is pir+, and is marked with an antibiotic resistance other than kanamycin and other than the donor strain.

to:

A significant benefit of this method is that, in theory, the user has wide latitude in the choice of enterobacterial recipient strains. This paper, as well as others11, use E. coli as a recipient strain, however the pJA1 plasmid has been used successfully with other enterobacterial recipient species such as Shigella flexneri6 and Salmonella enterica serovar Typhimurium strain SL134410. Theoretically, the γ origin of replication (oriR6Kγ) in pJA1 allows this plasmid to be maintained in a broad host range19, allowing that the recipient strain is pir-. The 300 base pair mob region from the RP4 plasmid in pJA1 allows conjugative transfer of this plasmid to a wide range of gram negative bacterial strains19. Simply put, this method could theoretically be used with a variety of recipient strains, as long as several conditions are met: the strain is pir-, and is marked with an antibiotic resistance other than kanamycin and other than the donor strain.

The References section was updated from:

  1. Blattner, F. R., Plunkett, G., et al. The Complete Genome Sequence of Escherichia coli K-12. Science. 277 (5331), 1453-1462 (1997).
  2. Alexeyev, M. F., & Shokolenko, I. N. Mini-Tn10 transposon derivatives for insertion mutagenesis and gene delivery into the chromosome of gram-negative bacteria. Gene. 160 (1), 59-62 (1995).
  3. Kvitko, B. H., Bruckbauer, S., et al. A simple method for construction of pir+ Enterobacterial hosts for maintenance of R6K replicon plasmids. BMC Res Notes. 5 (1), 157 (2012).
  4. Barrick, J. E., Yu, D. S., et al. Genome evolution and adaptation in a long-term experiment with Escherichia coli. Nature. 461 (7268), 1243-1247 (2009).
  5. Jacobs, M. A., Alwood, A., et al. Comprehensive transposon mutant library of Pseudomonas aeruginosa. PNAS. 100 (24), 14339-14344 (2003).
  6. van Opijnen, T., & Camilli, A. Transposon insertion sequencing: a new tool for systems-level analysis of microorganisms. Nature Rev Microbiol. 11 (7), 435-442 (2013).

to:

  1. Blattner, F. R., Plunkett, G., et al. The Complete Genome Sequence of Escherichia coli K-12. Science. 277 (5331), 1453-1462 (1997).
  2. Alexeyev, M. F., & Shokolenko, I. N. Mini-Tn10 transposon derivatives for insertion mutagenesis and gene delivery into the chromosome of gram-negative bacteria. Gene. 160 (1), 59-62 (1995).
  3. Barrick, J. E., Yu, D. S., et al. Genome evolution and adaptation in a long-term experiment with Escherichia coli. Nature. 461 (7268), 1243-1247 (2009).
  4. Jacobs, M. A., Alwood, A., et al. Comprehensive transposon mutant library of Pseudomonas aeruginosa. PNAS. 100 (24), 14339-14344 (2003).
  5. van Opijnen, T., & Camilli, A. Transposon insertion sequencing: a new tool for systems-level analysis of microorganisms. Nature Rev Microbiol. 11 (7), 435-442 (2013).

Disclosures

No conflicts of interest declared.

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