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Species:
Genetic Insert:
Vector Backbone Description: Vector Backbone:none; Vector Types:; Bacterial Resistance:None
References:
Comments: Genotype = ΔompF
Precursor strain = BL21Gold(DE3) [genotype F- ompT hsdS(rB– mB– ) dcm+ Tetr gal λ(DE3) endA Hte]
Supplemental document contains a list of genotypes and PCR primers used for verification of each knocked-out gene
Proper citation: RRID:Addgene_102259 Copy
Species:
Genetic Insert:
Vector Backbone Description: Vector Backbone:none; Vector Types:; Bacterial Resistance:None
References:
Comments: Genotype = ΔompA ΔompC ΔompF
Precursor strain = BL21Gold(DE3) [genotype F- ompT hsdS(rB– mB– ) dcm+ Tetr gal λ(DE3) endA Hte]
Supplemental document contains a list of genotypes and PCR primers used for verification of each knocked-out gene
Proper citation: RRID:Addgene_102268 Copy
Species:
Genetic Insert:
Vector Backbone Description: Vector Backbone:none; Vector Types:; Bacterial Resistance:None
References:
Comments: Genotype = ΔlamB ΔompC ΔompF
Precursor strain = BL21Gold(DE3) [genotype F- ompT hsdS(rB– mB– ) dcm+ Tetr gal λ(DE3) endA Hte]
Supplemental document contains a list of genotypes and PCR primers used for verification of each knocked-out gene
Proper citation: RRID:Addgene_102269 Copy
Species:
Genetic Insert: none
Vector Backbone Description: Vector Backbone:none; Vector Types:; Bacterial Resistance:None
References:
Comments: This work is supported in part by JSPS-NSF International Collaborations in Chemistry (ICC) research grant.
Genotype= thrC ilvA
Precursor strain = RF2
Modified from the parent Escherichia coli BL21(DE3) strain
Selective amino acid labeling (and/or requirement) = Thr, Ile
EH1 requires L-Thr and L-Ile for growth in M63 minimal medium
Please visit the following links for additional details on this strain and selective amino acid labeling-
http://www2.nms.ac.jp/fesworld/EcoliStrains.html
http://www2.nms.ac.jp/fesworld/EcoliStrainsSuppl.html
Note that these strains are NOT competent cells and one needs to make them competent before use.
Supplemental documents contain a list of PCR primers used for verification of each knocked-out gene as well as an image showing PCR results for this strain.
Supporting References:
Lin, M. T., Fukazawa, R., Miyajima-Nakano, Y., Matsushita, S., Choi, S. K., Iwasaki, T., and Gennis, R. B. (2015) Escherichia coliauxotroph host strains for amino acid-selective isotope labeling of recombinant proteins. Methods Enzymol. (Isotope Labeling of Biomolecules - Labeling Methods), 565, 45-66.
Iwasaki, T., Fukazawa, R., Miyajima-Nakano, Y., Baldansuren, A., Matsushita, S., Lin, M. T., Gennis, R. B., Hasegawa, K., Kumasaka, T., and Dikanov, S. A. (2012) Dissection of hydrogen bond interaction network around an iron-sulfur cluster by site-specific isotope labeling of hyperthermophilic archaeal Rieske-type ferredoxin. J. Am. Chem. Soc. 134, 19731-19738.
Lin, M. T., Sperling, L. J., Frericks Schmidt, H. L., Tang, M., Samoilova, R. I., Kumasaka, T., Iwasaki, T., Dikanov, S. A., Rienstra, C. M., and Gennis, R. B. (2011) A rapid and robust method for selective isotope labeling of proteins. Methods 55, 370-378.
Proper citation: RRID:Addgene_102804 Copy
Species:
Genetic Insert: none
Vector Backbone Description: Vector Backbone:none; Vector Types:; Bacterial Resistance:None
References:
Comments: This work is supported in part by JSPS-NSF International Collaborations in Chemistry (ICC) research grant.
Genotype= aspC tyrB trpA trpB glyA serB cysE
Precursor strain = RF15
Modified from the parent Escherichia coli BL21(DE3) strain
Selective amino acid labeling (and/or requirement) = Asp, Tyr, Trp, (Phe), Gly, Ser, Cys++++++, Ala++++++
++++++ RF16 grows slowly in the LB medium, does NOT grow in M63 minimal medium in the presence of L-Asp, L-Tyr, L-Trp, L-Gly, L-Ser plus L-Cys. RF16 (but not RF15) grows very slowly in M63 minimal medium in the presence of L-Asp, L-Tyr, L-Trp, L-Gly, L-Ser, L-Cys plus L-Ala.
Please visit the following links for additional details on this strain and selective amino acid labeling-
http://www2.nms.ac.jp/fesworld/EcoliStrains.html
http://www2.nms.ac.jp/fesworld/EcoliStrainsSuppl.html
Note that these strains are NOT competent cells and one needs to make them competent before use.
Supplemental documents contain a list of PCR primers used for verification of each knocked-out gene as well as an image showing PCR results for this strain.
Supporting References:
Lin, M. T., Fukazawa, R., Miyajima-Nakano, Y., Matsushita, S., Choi, S. K., Iwasaki, T., and Gennis, R. B. (2015) Escherichia coliauxotroph host strains for amino acid-selective isotope labeling of recombinant proteins. Methods Enzymol. (Isotope Labeling of Biomolecules - Labeling Methods), 565, 45-66.
Iwasaki, T., Fukazawa, R., Miyajima-Nakano, Y., Baldansuren, A., Matsushita, S., Lin, M. T., Gennis, R. B., Hasegawa, K., Kumasaka, T., and Dikanov, S. A. (2012) Dissection of hydrogen bond interaction network around an iron-sulfur cluster by site-specific isotope labeling of hyperthermophilic archaeal Rieske-type ferredoxin. J. Am. Chem. Soc. 134, 19731-19738.
Lin, M. T., Sperling, L. J., Frericks Schmidt, H. L., Tang, M., Samoilova, R. I., Kumasaka, T., Iwasaki, T., Dikanov, S. A., Rienstra, C. M., and Gennis, R. B. (2011) A rapid and robust method for selective isotope labeling of proteins. Methods 55, 370-378.
Proper citation: RRID:Addgene_102800 Copy
Species:
Genetic Insert: none
Vector Backbone Description: Vector Backbone:none; Vector Types:; Bacterial Resistance:None
References:
Comments: This work is supported in part by JSPS-NSF International Collaborations in Chemistry (ICC) research grant.
Genotype= aspC tyrB ilvE
Precursor strain = RF4
Modified from the parent Escherichia coli BL21(DE3) strain
Selective amino acid labeling (and/or requirement) = Asp, Tyr, Phe, Ile, Leu
Please note- RF17 strain has knockouts in aspC, tyrB, and ilvE genes and requires the presence of L-Asp, L-Tyr, L-Phe, L-Ile plus L-Leu for (slow) growth in M63 minimal medium.
Please visit the following links for additional details on this strain and selective amino acid labeling-
http://www2.nms.ac.jp/fesworld/EcoliStrains.html
http://www2.nms.ac.jp/fesworld/EcoliStrainsSuppl.html
Note that these strains are NOT competent cells and one needs to make them competent before use.
Supplemental documents contain a list of PCR primers used for verification of each knocked-out gene as well as an image showing PCR results for this strain.
Supporting References:
Lin, M. T., Fukazawa, R., Miyajima-Nakano, Y., Matsushita, S., Choi, S. K., Iwasaki, T., and Gennis, R. B. (2015) Escherichia coliauxotroph host strains for amino acid-selective isotope labeling of recombinant proteins. Methods Enzymol. (Isotope Labeling of Biomolecules - Labeling Methods), 565, 45-66.
Iwasaki, T., Fukazawa, R., Miyajima-Nakano, Y., Baldansuren, A., Matsushita, S., Lin, M. T., Gennis, R. B., Hasegawa, K., Kumasaka, T., and Dikanov, S. A. (2012) Dissection of hydrogen bond interaction network around an iron-sulfur cluster by site-specific isotope labeling of hyperthermophilic archaeal Rieske-type ferredoxin. J. Am. Chem. Soc. 134, 19731-19738.
Lin, M. T., Sperling, L. J., Frericks Schmidt, H. L., Tang, M., Samoilova, R. I., Kumasaka, T., Iwasaki, T., Dikanov, S. A., Rienstra, C. M., and Gennis, R. B. (2011) A rapid and robust method for selective isotope labeling of proteins. Methods 55, 370-378.
Proper citation: RRID:Addgene_102801 Copy
Species:
Genetic Insert: MG1655 Z1 malE clpA
Vector Backbone Description: Vector Backbone:n/a; Vector Types:; Bacterial Resistance:None
References:
Comments: F-, lambda-, rph-1, laciq, PN25-tetR, SpR, malE-, clpA-
Proper citation: RRID:Addgene_65918 Copy
Species:
Genetic Insert: MG1655 Z1 malE aat
Vector Backbone Description: Vector Backbone:n/a; Vector Types:; Bacterial Resistance:None
References:
Comments: F-, lambda-, rph-1, laciq, PN25-tetR, SpR, malE-, aat-
Proper citation: RRID:Addgene_65917 Copy
Species:
Genetic Insert:
Vector Backbone Description: Backbone Size:4600000; Vector Backbone:E. coli K-12 MG1655 genome; Vector Types:; Bacterial Resistance:None
References:
Comments: This strain carries the DE3 lysogen.
Proper citation: RRID:Addgene_61440 Copy
Species:
Genetic Insert: none
Vector Backbone Description: Vector Backbone:none; Vector Types:; Bacterial Resistance:None
References:
Comments: Genotype = tyrA
Precursor strain = C43(DE3)
modified from the parent Escherichia coli C43(DE3) strain
Selective amino acid labeling (and/or requirement) = (Tyr)
Each target gene was deleted from the chromosome of C43 (DE3) E. coli strain using λ-Red recombination system.
Supplemental documents contain a list of PCR primers used for verification of each knocked-out gene as well as an image showing PCR results for this strain.
Proper citation: RRID:Addgene_61911 Copy
Species:
Genetic Insert: none
Vector Backbone Description: Vector Backbone:none; Vector Types:; Bacterial Resistance:None
References:
Comments: Genotype = glnA
Precursor strain = C43(DE3)
modified from the parent Escherichia coli C43(DE3) strain
Selective amino acid labeling (and/or requirement) = Gln
Each target gene was deleted from the chromosome of C43 (DE3) E. coli strain using λ-Red recombination system.
Supplemental documents contain a list of PCR primers used for verification of each knocked-out gene as well as an image showing PCR results for this strain.
Proper citation: RRID:Addgene_61912 Copy
Species:
Genetic Insert: none
Vector Backbone Description: Vector Backbone:none; Vector Types:; Bacterial Resistance:None
References:
Comments: Genotype = cyo ilvE avtA aspC hisG metA
Precursor strain = ML23
modified from the parent Escherichia coli C43(DE3) strain
Selective amino acid labeling (and/or requirement) = (Ala#), Ile, Leu*, Tyr#, Val, His, Met
*In the presence of 0.4-1 mM Tyr, tyrB is repressed and Leu is required for growth in minimal medium. This strategy can only be applicable for a short-term cultivation but not suitable for a long-term cultivation for heterologous expression of foreign genes [Methods 55, 370-378 (2011)].
#In the presence of 0.4-1 mM Tyr, tyrB is repressed and Tyr is required for growth in minimal medium. Under these conditions, this strain can also be used for selective labeling of the input Tyr and/or Ala label(s), although minor diffusion of the input label(s) can occur. Note that this strategy can only be applicable for a short-term cultivation but not suitable for a long-term cultivation for heterologous expression of foreign geness [Methods 55, 370-378 (2011)].
Each target gene was deleted from the chromosome of C43 (DE3) E. coli strain using λ-Red recombination system.
Supplemental documents contain a list of PCR primers used for verification of each knocked-out gene as well as an image showing PCR results for this strain.
Proper citation: RRID:Addgene_61919 Copy
Species:
Genetic Insert: none
Vector Backbone Description: Vector Backbone:none; Vector Types:; Bacterial Resistance:None
References:
Comments: Genotype = cyo ilvE avtA aspC hisG asnA
Precursor strain = ML23
modified from the parent Escherichia coli C43(DE3) strain
Selective amino acid labeling (and/or requirement) = (Ala#), Ile, Leu*, Tyr#, Val, His
*In the presence of 0.4-1 mM Tyr, tyrB is repressed and Leu is required for growth in minimal medium. This strategy can only be applicable for a short-term cultivation but not suitable for a long-term cultivation for heterologous expression of foreign genes [Methods 55, 370-378 (2011)].
#In the presence of 0.4-1 mM Tyr, tyrB is repressed and Tyr is required for growth in minimal medium. Under these conditions, this strain can also be used for selective labeling of the input Tyr and/or Ala label(s), although minor diffusion of the input label(s) can occur. Note that this strategy can only be applicable for a short-term cultivation but not suitable for a long-term cultivation for heterologous expression of foreign geness [Methods 55, 370-378 (2011)].
Each target gene was deleted from the chromosome of C43 (DE3) E. coli strain using λ-Red recombination system.
Supplemental documents contain a list of PCR primers used for verification of each knocked-out gene as well as an image showing PCR results for this strain.
Proper citation: RRID:Addgene_61915 Copy
Species:
Genetic Insert: none
Vector Backbone Description: Vector Backbone:none; Vector Types:; Bacterial Resistance:None
References:
Comments: Genotype = cyo ilvE avtA aspC hisG asnA asnB
Precursor strain = ML24
modified from the parent Escherichia coli C43(DE3) strain
Selective amino acid labeling (and/or requirement) = (Ala#), Ile, Leu*, Tyr#, Val, His, Asn
*In the presence of 0.4-1 mM Tyr, tyrB is repressed and Leu is required for growth in minimal medium. This strategy can only be applicable for a short-term cultivation but not suitable for a long-term cultivation for heterologous expression of foreign genes [Methods 55, 370-378 (2011)].
#In the presence of 0.4-1 mM Tyr, tyrB is repressed and Tyr is required for growth in minimal medium. Under these conditions, this strain can also be used for selective labeling of the input Tyr and/or Ala label(s), although minor diffusion of the input label(s) can occur. Note that this strategy can only be applicable for a short-term cultivation but not suitable for a long-term cultivation for heterologous expression of foreign geness [Methods 55, 370-378 (2011)].
Each target gene was deleted from the chromosome of C43 (DE3) E. coli strain using λ-Red recombination system.
Supplemental documents contain a list of PCR primers used for verification of each knocked-out gene as well as an image showing PCR results for this strain.
Proper citation: RRID:Addgene_61916 Copy
Species:
Genetic Insert: none
Vector Backbone Description: Vector Backbone:none; Vector Types:; Bacterial Resistance:None
References:
Comments: Genotype = cyo ilvE avtA aspC hisG argH metA lysA thrC asnB
Precursor strain = ML41
modified from the parent Escherichia coli C43(DE3) strain
Selective amino acid labeling (and/or requirement) = (Ala#), Ile, Leu*, Tyr#, Val, His, Arg, Met, Lys, Thr
*In the presence of 0.4-1 mM Tyr, tyrB is repressed and Leu is required for growth in minimal medium. This strategy can only be applicable for a short-term cultivation but not suitable for a long-term cultivation for heterologous expression of foreign genes [Methods 55, 370-378 (2011)].
#In the presence of 0.4-1 mM Tyr, tyrB is repressed and Tyr is required for growth in minimal medium. Under these conditions, this strain can also be used for selective labeling of the input Tyr and/or Ala label(s), although minor diffusion of the input label(s) can occur. Note that this strategy can only be applicable for a short-term cultivation but not suitable for a long-term cultivation for heterologous expression of foreign geness [Methods 55, 370-378 (2011)].
Each target gene was deleted from the chromosome of C43 (DE3) E. coli strain using λ-Red recombination system.
Supplemental documents contain a list of PCR primers used for verification of each knocked-out gene as well as an image showing PCR results for this strain.
Proper citation: RRID:Addgene_61921 Copy
Species:
Genetic Insert: none
Vector Backbone Description: Vector Backbone:none; Vector Types:; Bacterial Resistance:None
References:
Comments: Genotype = metA
Precursor strain = C43(DE3)
Selective amino acid labeling (and/or requirement) = Met
Each target gene was deleted from the chromosome of C43 (DE3) E. coli strain using λ-Red recombination system.
Supplemental documents contain a list of PCR primers used for verification of each knocked-out gene as well as an image showing PCR results for this strain.
This work is supported in part by JSPS-NSF International Collaborations in Chemistry (ICC) research grant.
Proper citation: RRID:Addgene_61961 Copy
Species:
Genetic Insert: none
Vector Backbone Description: Vector Backbone:none; Vector Types:; Bacterial Resistance:None
References:
Comments: Genotype = thrC
Precursor strain = BL21 CodonPlus (DE3)-RIL
Selective amino acid labeling (and/or requirement) = Thr
Each target gene was deleted from the chromosome of BL21 (DE3) E. coli strain using λ-Red recombination system.
Supplemental documents contain a list of PCR primers used for verification of each knocked-out gene as well as an image showing PCR results for this strain.
This work is supported in part by JSPS-NSF International Collaborations in Chemistry (ICC) research grant.
Proper citation: RRID:Addgene_62070 Copy
Species:
Genetic Insert: none
Vector Backbone Description: Vector Backbone:none; Vector Types:; Bacterial Resistance:None
References:
Comments: Genotype = trpA trpB
Precursor strain = BL21 CodonPlus (DE3)-RIL
Selective amino acid labeling (and/or requirement) = Trp##
##RF12 strain requires Trp for growth in M63 minimal medium. For effective Trp labeling of a recombinant protein, the presence of 0.4-1 mM Tyr in growth medium to repress the tyrB gene is recommended. This strategy can only be applicable for a short-term cultivation but not suitable for a long-term cultivation for heterologous expression of foreign genes [Methods 55, 370-378 (2011)].
Each target gene was deleted from the chromosome of BL21 (DE3) E. coli strain using λ-Red recombination system.
Supplemental documents contain a list of PCR primers used for verification of each knocked-out gene as well as an image showing PCR results for this strain.
This work is supported in part by JSPS-NSF International Collaborations in Chemistry (ICC) research grant.
Proper citation: RRID:Addgene_62077 Copy
Species:
Genetic Insert: none
Vector Backbone Description: Vector Backbone:none; Vector Types:; Bacterial Resistance:None
References:
Comments: Genotype = aspC tyrB hisG
Precursor strain = RF4
modified from the parent Escherichia coli BL21 CodonPlus (DE3)-RIL
Selective amino acid labeling (and/or requirement) = Asp++++, Tyr, (Phe), His
++++RF4, RF5, and RF13 strains have knockouts in both aspC and tyrB genes and therefore require the presence of L-Asp (but not L-Glu) for growth in M63 minimal medium. These strains can be used for selective labeling of selected aromatic amino acid(s) such as Tyr (RF4, RF5, RF13) and/or Trp (RF13). We have NOT tested if these strains could also be used for selective Asp labeling studies (under investigation).
Each target gene was deleted from the chromosome of BL21 (DE3) E. coli strain using λ-Red recombination system.
Supplemental documents contain a list of PCR primers used for verification of each knocked-out gene as well as an image showing PCR results for this strain.
This work is supported in part by JSPS-NSF International Collaborations in Chemistry (ICC) research grant.
Proper citation: RRID:Addgene_62073 Copy
Species:
Genetic Insert: none
Vector Backbone Description: Vector Backbone:none; Vector Types:; Bacterial Resistance:None
References:
Comments: Genotype = lysA
Precursor strain = BL21 CodonPlus (DE3)-RIL
Selective amino acid labeling (and/or requirement) = Lys
Each target gene was deleted from the chromosome of BL21 (DE3) E. coli strain using λ-Red recombination system.
Supplemental documents contain a list of PCR primers used for verification of each knocked-out gene as well as an image showing PCR results for this strain.
This work is supported in part by JSPS-NSF International Collaborations in Chemistry (ICC) research grant.
Proper citation: RRID:Addgene_62076 Copy
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