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So far, only 5,000 genes have been mutated, and the International Mouse Mutagenesis Consortium aims to generate strains with mutations in the remaining 25,000.
RAPD was used to generate strain-specific markers.
The C. efficiens NBRC 100395 (YS-314) vanillate O-demethylase gene (vanABCE; CE0634 CE0635) was expressed in C. glutamicum ATCC 21420 to generate strain FVan.
A more favorable effect of this recombinant strain on sorghum root growth and development suggests that metabolic engineering could be used to generate strains with improved PGP function.
We introduced these plasmids into E. coli DH1 [11] to generate strain B32 (Tables 1 and 2).
Contacts involving many beta-branched hydrophobic residues in the crowded protein core may generate strain in helices, destabilizing the structure.
The rpoE gene was deleted first, using the method of Datsenko and Wanner [63] to generate strain SEA4041.
The resulting strains (MinC, SB080 and MinD, SB076) were subsequently transformed with minJ tet genomic DNA to generate strains SB081 (MinC+ ΔminJ) and SB077 (MinD+ ΔminJ).
To introduce a CYS4(1–353)-13MYC::Kallelellele in the standard BY4742 (MATα; his3Δ; leu2Δ; lystrainra3Δ) strain, we followed a single-step PCR replacement protocol as described [37], to generate strain SCMSP179.
The nadB::Tn10 insertion from SEA2000 was then moved into the rpoE::kan strain by P1 transduction, selecting for resistance to Kan and Tet to generate strain SEA4114.
The resulting plasmid (R995+SPI-19) was used to complement our S. Gallinarum SPI-19 deletion mutant in trans to generate strain ΔSPI-19/R995+SPI-19.
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