Category:Strain:BW25113
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<protect>
| Name |
BW25113 |
|---|---|
| Synonyms | |
| Genotype |
F- DE(araD-araB)567 lacZ4787(del)::rrnB-3 LAM- rph-1 DE(rhaD-rhaB)568 hsdR514 |
| Phenotype | |
| Plasmids | |
| Phage | |
| Sources | |
| Construction |
see description below |
| References | |
</protect>
Notes
BW25113 was used to make the Keio Collection of single-gene knockouts. [2] [1] BW25113 is available from the Coli Genetics Stock Center: CGSC#7636 The genotype for BW25113 in Datsenko et al. [1] said the strain is lacIQ, but it was later shown to be lacI+ Baba et al. (Supplementary Table 1)[2].
According to the CGSC, lacZ4787(del) has 4 tandem copies of the rrnB transcriptional terminator inserted by gene replacement into the region extending from near the SacII site near the N-terminus of lacZ through the promoter.
BW25113 is a derivative of E. coli K-12 strain BD792, which has the genotype F- LAM- rpoS396(Am) rph-1.
The steps in the construction of BW25113 are described below. This information is from Supplementary Table 1 of Baba et al.([2]). At several steps of the strain construction, mutant alleles were constructed in vitro and recombined into the genome using oriRR6Kgamma plasmids, which cannot replicate in a strain that lacks the R6K pir gene ([3]). In the absence of selection of the antibiotic resistance marker carried on the plasmid, homologous recombination can occur that will either restore the original genotype or result in allele replacement.
| Step | Strain | Genotype | Construction | Reference |
|---|---|---|---|---|
|
ii) |
BW21597 |
F- LAM- rpoS396(Am) DEphoR574 rph-1 |
BD792 was made TcR with pAH14 then TcS Pho- derivatives were isolated.[4] |
unpublished |
|
iii) |
BW21652 |
F- LAM- rpoS396(Am) DEphoR574 DE(creABCD)154 rph-1 |
BW21597 was made TcR with pSK26 and then TcS Cre- derivatives were isolated. [3] |
unpublished |
|
iv) |
BW23186 |
F- LAM- rpoS396(Am) DEphoR574 DE(creABCD)154 DE(araBAD)567 rph-1 |
BW21652 was made TcR with pAH54 and then TcS Ara- derivatives were isolated. ([5]) |
unpublished |
|
v) |
BW23599 |
F- LAM- rpoS396(Am) rrnB3 DElacZ4787 proC::Tn5-132 DE(creABCD)154 DE(araBAD)567 rph-1 |
BW23186 was transduced to TcR Pro- Lac- Pho+ with P1kc grown on BW22773[5] |
unpublished |
|
vi) |
BW23660 |
F- LAM- rpoS396(Am) rrnB3 DElacZ4787 DE(phoBR)580 DE(creABCD)154 DE(araBAD)567 rph-1 |
BW23599 was transduced to Pro+ Pho- with P1kc grown on BW23316. |
Haldimann et al. 1997[4] |
|
vii) |
BW24200 |
F- LAM- rpoS396(Am) rrnB3 DElacZ4787 DE(phoBR)580 DE(creABCD)154 DE(araBAD)567 metF159(Am) zii-510::Tn10 rph-1 |
BW23660 was transduced to TcR Met- with P1kc grown on JW383 (Haldimann et al., 1997) |
unpublished |
|
viii) |
BW24201 |
F- LAM- rpoS396(Am) rrnB3 DElacZ4787 DE(phoBR)580 DE(creABCD)154 DE(araBAD)567 DE(rhaBAD)568 rph-1 |
BW24200 was transduced to Met+ TcS Rha- with P1kc grown on BW22875[5] |
unpublished |
|
ix) |
BW24310 |
F- LAM- rrnB3 DElacZ4787 DE(phoBR)580 DE(creABCD)154 DE(araBAD)567 DE(rhaBAD)568 cysC95::Tn10 rpoS::kan rph-1 |
BW24201 was transduced to TcR KmR with P1kc grown on ZK1001[5] |
Haldimann and Wanner, 2001[6] |
|
x) |
BW24320a |
F- LAM- rrnB3 DElacZ4787 DE(phoBR)580 DE(creABCD)154 DE(araBAD)567 DE(rhaBAD)568 rph-1 |
BW24310 was transduced to Cys+ KmS with P1kc grown on MG1655 (Wanner and Boline, 1990) aBW24320 and its descendants carry the same pseudorevertant rpoS(Q33) allele as MG1655 (Hayashi et al., 2005) |
Haldimann and Wanner, 2001[6] |
|
xi) |
BW24353 |
F- LAM- rrnB3 DElacZ4787 DE(phoBR)580 DE(zaa trpR creABCD arcA thr)104::Tn5-132 DE(araBAD)567 DE(rhaBAD)568 rph-1 |
BW24320 was transduced to TcR Thr- with P1kc grown on BW12070 (Fisher et al., 1995) |
unpublished |
|
xii) |
BW24476 |
F- LAM- rrnB3 DElacZ4787 DE(phoBR)580 hsdR514 DE(araBAD)567 DE(rhaBAD)568 rph-1 |
BW24353 was transduced to Thr+ HsdR- with P1kc grown on BW24447[7] |
Stern et al., 1999[7] |
|
xiii) |
BW25083 |
F- LAM- rrnB3 DElacZ4787 proC::Tn5-132 hsdR514 DE(araBAD)567 DE(rhaBAD)568 rph-1 |
BW24476 was transduced to TcR Pro- Pho+ with P1kc grown on BW22773[5] |
Haldimann and Wanner, 2001[6] |
|
xiv) |
BW25113 |
F- LAM- rrnB3 DElacZ4787 hsdR514 DE(araBAD)567 DE(rhaBAD)568 rph-1 |
BW25083 was transduced to Pro+ with P1kc grown on BW24321[6] |
|
References
See Help:References for how to manage references in EcoliWiki.
- ↑ 1.0 1.1 1.2 1.3 Datsenko, KA & Wanner, BL (2000) One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products. Proc. Natl. Acad. Sci. U.S.A. 97 6640-5 PubMed
- ↑ 2.0 2.1 2.2 2.3 Baba, T et al. (2006) Construction of Escherichia coli K-12 in-frame, single-gene knockout mutants: the Keio collection. Mol. Syst. Biol. 2 2006.0008 PubMed
- ↑ 3.0 3.1 Metcalf, WW et al. (1996) Conditionally replicative and conjugative plasmids carrying lacZ alpha for cloning, mutagenesis, and allele replacement in bacteria. Plasmid 35 1-13 PubMed
- ↑ 4.0 4.1 Haldimann, A et al. (1997) Transcriptional regulation of the Enterococcus faecium BM4147 vancomycin resistance gene cluster by the VanS-VanR two-component regulatory system in Escherichia coli K-12. J. Bacteriol. 179 5903-13 PubMed
- ↑ 5.0 5.1 5.2 5.3 5.4 Haldimann, A et al. (1998) Use of new methods for construction of tightly regulated arabinose and rhamnose promoter fusions in studies of the Escherichia coli phosphate regulon. J. Bacteriol. 180 1277-86 PubMed
- ↑ 6.0 6.1 6.2 6.3 Haldimann, A & Wanner, BL (2001) Conditional-replication, integration, excision, and retrieval plasmid-host systems for gene structure-function studies of bacteria. J. Bacteriol. 183 6384-93 PubMed
- ↑ 7.0 7.1 Stern, RJ et al. (1999) Conversion of dTDP-4-keto-6-deoxyglucose to free dTDP-4-keto-rhamnose by the rmIC gene products of Escherichia coli and Mycobacterium tuberculosis. Microbiology (Reading, Engl.) 145 ( Pt 3) 663-71 PubMed
- ↑ Lessard, IA et al. (1998) Homologs of the vancomycin resistance D-Ala-D-Ala dipeptidase VanX in Streptomyces toyocaensis, Escherichia coli and Synechocystis: attributes of catalytic efficiency, stereoselectivity and regulation with implications for function. Chem. Biol. 5 489-504 PubMed
