Project description:The WP3 strains was cultured at 20°C/0.1 MPa, and then was incubated at high hydrostatic pressure (HHP, 50 MPa) for 120 min. The transcriptional profiles after and before HHP shock were compared.
Project description:The WP3 strains was cultured at 20°C/0.1 MPa, and then was incubated at high hydrostatic pressure (HHP, 50 MPa) for 30 min. The transcriptional profiles after and before HHP shock were compared.
Project description:Most of the Shewanella species contain two periplasmic nitrate reductases (NAP-? and NAP-?), which is a unique feature of this genus. In the present study, the physiological function and evolutionary relationship of the two NAP systems were studied in the deep-sea bacterium Shewanella piezotolerans WP3. Both of the WP3 nap gene clusters: nap-? (napD1A1B1C) and nap-? (napD2A2B2) were shown to be involved in nitrate respiration. Phylogenetic analyses suggest that NAP-? originated earlier than NAP-?. Tetraheme cytochromes NapC and CymA were found to be the major electron deliver proteins, and CymA also served as a sole electron transporter towards nitrite reductase. Interestingly, a ?napA2 mutant with the single functional NAP-? system showed better growth than the wild-type strain, when grown in nitrate medium, and it had a selective advantage to the wild-type strain. On the basis of these results, we proposed the evolution direction of nitrate respiration system in Shewanella: from a single NAP-? to NAP-? and NAP-? both, followed by the evolution to a single NAP-?. Moreover, the data presented here will be very useful for the designed engineering of Shewanella for more efficient respiring capabilities for environmental bioremediation.
Project description:The cAMP binding domain of crp gene was deleted from WP3 genome to construct the WP3ΔcrpD1 strain. The two strains were cultured at 20°C and the transcriptional profiles were compared.
Project description:The DNA binding domain of crp gene was deleted from WP3 genome to construct the WP3ΔcrpD2 strain. The two strains were cultured at 20°C and the transcriptional profiles were compared.
Project description:The cAMP receptor protein (CRP) is a conserved regulator in bacteria and involved in regulation of energy metabolism, such as glucose, galactose, and citrate (Green et al., 2014 [1]). As an important catabolite activator protein, it has been well characterized in model microorganism such as Escherichia coli. However, our understanding of the roles of CRP in deep-sea bacteria is rather limited. To indentify the function of CRP, we performed whole genome transcriptional profiling using a custom designed microarray which contains 95% open reading frames of Shewanella piezotolerans WP3, which was isolated from West Pacific sediment at a depth of 1914 m (Xiao et al., 2007 [2]; Wang et al., 2008 [3]). Here we describe the experimental procedures and methods in detail to reproduce the results (available at Gene Expression Omnibus database under GSE67731 and GSE67732) and provide resource to be employed for comparative analyses of CRP regulon and the regulatory network of anaerobic respiration in microorganisms which inhabited in different environments, and thus broaden our understanding of mechanism of bacteria against various environment stresses.
Project description:The hns gene was deleted from WP3 genome to construct the WP3Δhns strain. The two strains were cultured at 20°C and the transcriptional profiles were compared.
Project description:The WP3 strains was cultured at 20°C and heat shocked at 40°C for 90 min. The transcriptional profiles after and before heat shock were compared.
Project description:The WP3 strains was cultured at 20°C and heat shocked at 40°C for 60 min. The transcriptional profiles after and before heat shock were compared.