The primary and secondary structure analyses were performed NVP-BKM120 purchase using the protparam tool on ExPASy server (Bairoch et al., 2005) and psipred (Jones,

1999), respectively. The tertiary structure of CspD from Ant5-2 was generated by the modeller software from hhpred alignments on HHpred servers (Soding et al., 2005; Eswar et al., 2006). The significance of the protein structure similarity was measured by TM-score calculated by T-align, a more sensitive method than root-mean-square-deviation (Zhang & Skolnick, 2005). The protein–protein docking was performed using the hex 5.1 software according to its manual (Ritchie & Venkatraman, 2010). During the initial 4 h, Ant5-2 cultures exhibited faster growth rates at 22 and 37 °C than at 15 °C (Fig. 1). Within 20 h of incubation at 15, 22 and 37 °C, the cultures grew exponentially and reached stationary phase. However, the culture at 37 °C exhibited a decline in growth after 48 h of incubation. In contrast, the cultures maintained at −1 and 4 °C did not show

any significant cell proliferation for 24 and 4 h, respectively. Thereafter, the culture at 4 °C exhibited exponential growth and reached stationary phase within 72 h. A slow but steady exponential growth of the culture at −1 °C was noticed after incubation for 24 h. After one freeze–thaw cycle, increased survival was observed when Ant5-2 was exposed to 4 °C before freezing. The cultures transferred to 4 °C showed 94.1% survival compared with the 48.9% survival of cultures incubated http://www.selleckchem.com/products/Tigecycline.html at 22 °C (Supporting Progesterone Information, Fig. S1). The autoradiogram exhibited expression of a ∼7.28-kDa Csp

(CspD) at all temperatures (Fig. S2). The immunoblot results showed that the expression of CspD in Ant5-2 was both time- and growth phase-dependent (Fig. 2a). Its expression increased at 37 °C and UVC exposure (Fig. 2b and c). A 204-bp DNA fragment encompassing the entire ORF of the cspD gene (accession no. HQ873479) was PCR amplified. The deduced amino acid sequence exhibited 100% identity with the cold shock transcription regulator of J. lividum DSM 1522 and >98% sequence identity with the RNA chaperone, transcription antiterminator of Herminiimonas arsenicoxydans and with Csps from different bacteria belonging to class Betaproteobacteria (Figs 3 and Fig. S3). The CspD from Ant5-2 showed highest identity and similarity to E. coli CspE (67/83%) and E. coli CspD (56/74%) when compared with Csps from E. coli, B. subtilis and Pseudomonas sp. 30/3 (Fig. 3). PCR amplification of the cspA family of genes in Ant5-2 genomic DNA using CSPU5 and CSPU3 universal primers (Francis and Stewart, 1997) resulted in negative outcome (Fig. S4).