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Host strain


E. coli strain for producing recombinant proteins incorporating synthetic amino acids

  • The host E. coli strains, RFzero-iy strain and B-95.deltaA strain that can produce recombinant proteins incorporating synthetic amino acids by reassignment of the UAG codon.
  • The RFzero-iy strains are designated to incorporate 3-iodotyrosine into produced proteins. For the incorporation, the strain is transformed with the expression plasmid in which a codon for the target tyrosine is replaced with UAG codon and cultured in 3-iodotyrosine containing medium (Mukai, T. et al., 2011). Two strains based on the BW25113 and BL-21(DE3) are available.
  • The B-95.deltaA strains can be assigned their UAG codon to synthetic amino acids by the introduction of plasmids carrying specific pair of UAG-reading tRNA and an aminoacyl-tRNA synthetase (aaRS) variant*. Increased productivity of sulfonated hirudin which is expected to improve the inhibition of blood coagulation was reported (Mukai, T. et al., 2015). The original and the growth improved derivative strains are available.
  • Reference
    Mukai, T. et al., Biochem. Biophys. Res. Commun. 411 (4): 757-761, 2011. [PMID 21782790]
    Mukai, T. et al., Sci. Rep. 5: 9699, 2015. [PMID 25982672]
    Incorporation of synthetic amino acids into proteins at specific sites

    RDB13711 B95. delta A Escherichia coli BL-21(DE3)-based host strain with no specific assignment of the UAG codon.
    RDB13712 B95. delta A delta fabR Escherichia coli BL21(DE3)-based host strain with no specific assignment of the UAG codon.
    RDB14427 BW25113-based RFzero-iy Escherichia coli host strain to produce non-natural amino acids containing proteins.
    RDB14428 BL21(DE3)-based RFzero-iy Escherichia coli host strain to produce non-natural amino acids containing proteins.
  • *Plamids that introduce synthetic amino acids are provided by RIKEN CLST.

E. coli auxotroph host strain used for selective isotope labeling

  • All ML strains werre engineered by Dr. Gennis research group at University of Illinois at Urbana-Champaign, U.S.A., and YM, MS, EH, and RF strains were engineered by Dr. Iwasaki research group at Nippon Medical School, Japan. This work was supported in part by the Internaitonal Collaborations in Chemistry Grant from JSPS and NSF, the Nagase Science and Technology Foundation Research Grant, and JSPS grants-in-aid.
    Catalog# Strain name Genotype Amino acid requirement and/or labeling
    RDB15927 ML2 cyo::kan ilvE; Ile, Leu.
    RDB15928 ML3 cyo::kan hisG His.
    RDB15929 ML6 cyo::kan ilvE avtA Ile, Leu, Val.
    RDB15930 ML8 cyo::kan argH Arg.
    RDB15931 ML12 cyo::kan ilvE avtA aspC (Ala), Ile, Leu, Tyr, Val.
    RDB15932 ML14 tyrA (Tyr).
    RDB15933 ML17 glnA Gln.
    RDB15934 ML21 tyrA hisG (Tyr), His.
    RDB15935 ML24 cyo ilvE avtA aspC hisG asnA (Ala), Ile, Leu, Tyr, Val, His.
    RDB15936 ML25 cyo ilvE avtA aspC hisG asnA asnB (Ala), Ile, Leu, Tyr, Val, His, Asn.
    RDB15937 ML26 cyo ilvE avtA aspC hisG argH (Ala), Ile, Leu, Tyr, Val, His, Arg.
    RDB15938 ML31 cyo ilvE avtA aspC hisG argH metA (Ala), Ile, Leu, Tyr, Val, His, Arg, Met.
    RDB15939 ML36 cyo ilvE avtA aspC hisG metA (Ala), Ile, Leu, Tyr, Val, His, Met.
    RDB15940 ML40 cyo ilvE avtA aspC hisG argH metA lysA (Ala), Ile, Leu, Tyr, Val, His, Arg, Met, Lys.
    RDB15941 ML42 cyo ilvE avtA aspC hisG argH metA lysA thrC asnB (Ala), Ile, Leu, Tyr, Val, His, Arg, Met, Lys, Thr.
    RDB15942 ML43 cyo ilvE avtA aspC hisG argH metA lysA thrC asnA asnB (Ala), Ile, Leu, Tyr, Val, His, Arg, Met, Lys, Thr, Asn.
    RDB15943 YM138 cysE Cys.
    RDB15944 MS1 cysE hisG Cys, His.
    RDB15952 RF11 metA Met.
    RDB15945 RF1 glyA Gly.
    RDB15946 RF2 thrC Thr.
    RDB15947 RF4 aspC tyrB Asp, Tyr, (Phe).
    RDB15948 RF5 aspC tyrB hisG Asp, Tyr, (Phe), His.
    RDB15949 RF6 proC Pro.
    RDB15950 RF8 asnA asnB Asn.
    RDB15951 RF10 lysA Lys.
    RDB15953 RF12 trpA trpB Trp.
    RDB15954 RF13 aspC tyrB trpA trpB Asp, Tyr, Trp, (Phe).
    RDB15955 RF15 aspC tyrB trpA trpB glyA serB Asp, Tyr, Trp, (Phe), Gly, Ser.
    RDB15956 RF16 aspC tyrB trpA trpB glyA serB cysE Asp, Tyr, Trp, (Phe), Gly, Ser, Cys, Ala.
    RDB15957 RF17 aspC tyrB ilvE Asp, Tyr, Phe, Ile, Leu.
    RDB15958 RF18 aspC tyrB ilvE avtA Asp, Tyr, Phe, Ile, Leu, Val.
    RDB15959 RF21 aspC tyrB ilvE avtA yfbQ(alaA) yfdZ(alaC) Asp, Tyr, Phe, Ile, Leu, Val.
    RDB15960 EH1 thrC ilvA Thr, Ile.
  • Please refer web site of Escherichia coli auxotrophic expression strains (by the depositor), Lin et al., Methods 55 (4): 370-378 (2011), Iwasaki et al., J. Am. Chem. Soc. 134 (48): 19731-19738 (2012), and/or Lin et al., Methods Enzymol. 565: 45-66 (2015).

 

Bacillus empty Backbone

RDB00919 pSTE12 Bacillus stearothermophilus shuttle vector
RDB00920 pSTE33 Bacillus stearothermophilus shuttle vector

Bacillus stearothermophilus bacterial host

RDB00139 K1041 Host strain of Bacillus stearothermophilus for pSTE33.

(GRP0028e 2018.06.01 T.M.)

2018.06.01