Table 1.
Identification of promoters in upstream regions of vir and icm/dot genes from X. euvesicatoria strain 85-10.
| Gene | Length of promoter sequence1 | BROM prediction2 | vir box3 | GFP synthesis4 | GFP fluorescence5 | |
|---|---|---|---|---|---|---|
| -10 region | -35 region | |||||
| virB2 | 234 | + | + | + | + | +/- |
| virB5 | 231 | + | + | + | + | + |
| virD4 | 242 | – | – | – | + | – |
| virG | 301 | – | – | – | + | + |
| dotA | 240 | + | + | + | + | +/- |
| dotD | 207 | + | + | + | + | +/- |
| icmL229 | 229 | – | – | – | – | – |
| icmL129 | 129 | + | + | + | + | + |
1 Putative promoter fragments were cloned upstream of the sfgfp gene for expression studies in X. euvesicatoria strain 85-10 or upstream of dTALE-2 in strain 85-10ΔxopQ for the analysis of in planta translocation of dTALE-2 into gfp-transgenic N. benthamiana plants.
2 The prediction program BROM (Softberry) http://www.softberry.com/berry.phtml?topic=bprom&group=programs&subgroup=gfindb ) predicts bacterial sigma70 promoter regions.
3 vir box elements (5’-TG(A/T)AA(C/T)-3’) are present in putative promoter fragments of vir and icm/dot genes from X. euvesicatoria. +, presence of putative vir box; -, absence of putative vir box.
4 GFP synthesis in X. euvesicatoria was detected by immunoblot analysis of total cell extracts from bacteria grown in liquid NYG cultures. +, detection of GFP by immunoblotting; -, no GFP detectable (see Figure 2 ).
5 GFP fluorescence was monitored in gfp-transgenic N. benthamiana plants after infection with X. euvesicatoria strain 85-10ΔxopQ expressing dTALE-2 under control of vir and icm/dot promoter fragments from respective expression constructs. +, GFP fluorescence; -, no GFP fluorescence detectable; +/-, reduced GFP fluorescence.