Fig. 4

ku70 disruption of wild-type M. thermophila ATCC 42464. a Pattern of the ku70 knockout cassette integrating into the chromosomes of M. thermophila ATCC 42464 via ectopic insertion or homologous recombination. b PCR analysis of ku70 deletion transformants with one primer (ku70KO-F) located in the bar gene cassette and the other (ku70KO-R) located in the downstream of 3′ flank in the genomic DNA (expected product of 1872 bp) and c Southern blotting analysis of ku70 mutants with XhoI digested genomic DNA and the probe amplified from the 3′ flank. Lines 1–11 in (b) and (c), genomic DNA from ku70 mutants (expected product of 5056 bp). WT, genomic DNA from wild-type as negative control (expected product of 1703 bp). Primers are represented by solid black arrows. Abbreviations: gDNA, genomic DNA; RB and LB, right and left border of T-DNA, respectively