Based on the RNA-seq results of the Desulfovibrio vulgaris biofilm cell vs. planktonic cell, we have found that dvu2956, a sigma-54 dependent transcriptional regulator gene, was significantly repressed in SRB biofilm cell. Later DVU2956 was successfully cloned and fused biofilm phase promotor Pdvu0304 into D. vulgaris, which makes highly expression during biofilm formation. The biofilm assay showed that this recombinant strain D. vulgaris/pVLT33-P0304-dvu2956 biofilm was inhibited by 70% (48 hr) when compared with empty plasmid control strain. In this work, to reveal the biofilm regulation network, comparative transcriptomic analysis between biofilm cell of D. vulgaris/ pVLT33-Pdvu0304-dvu2956 and empty plasmid control strains by RNA-seq was performed following the strategy of SRB RNA-seq. Two replicates of each sample was used.