Fig. 2.

Confirmation of the in-frame deletion mutant Δagl3 (Saci0423).

A. Gene deletion was confirmed by PCR using the outside primers against the flanking regions of agl3, DNA isolated from the background strain MW001 (lane 2), plasmid pSVA1225 used for the homologous recombination incorporating the up- and downstream region of deleted agl3 (lane 3), or the mutant lacking agl3 (lane 1).

B. RT-PCR confirms the deletion of the gene agl3 presumably encoding the UDP-sulfoquinovose. The cDNA (C) served as a template in PCR amplification using primers against the internal region of either the agl3 or aglB. In each case, PCR amplifications were also performed using genomic DNA (G) as a positive control and total RNA (R) as a negative control.