처리결과

Umsporulation can explain why downregulation of environmental information-processing proteins in the Cu2+ added medium does not inhibit ICP expression, given that sporulation is a mechanism used by B. thuringiensis to resist unfavorable environmental conditions. Reduces the expression of environmental information-processing proteins, and utilizes carbon and nitrogen from intracellular poly--hydroxybutyrate (PHB), lipids and proteins stored in the logarithmic phase during sporulation. The macromolecular and small molecular metabolism proteins were further sorted according to the metabolic phases in which they are involved (Fig. 7b, c; Additional files 2, 3, 4). Fold changes of subcategories were also computed (Fig. 8b, c). For macromolecular metabolism, a large majority of the proteins were involved in protein synthesis, degradation and modification (Fig. 7b; Additional file 1). The subcategory of glycan biosynthesis and metabolism proteins showed downregulation (Fig. 8b), reducing the expression of extracellular polysaccharides. More carbon sources can be used to synthesize intracellular PHB. Other subcategories showed no significant difference. For small molecular metabolism, proteins that take part in amino acid metabolism and central carbon metabolism comprised the largest subcategory (Fig. 7c; Additional file 1). Fold changes of subcategories in small molecular metabolism showed no significant difference (Fig. 8c).emPAI semiquantitative comparative analysissubunit beta (ATPS), and protein PrkA. For the ICP, emPAI semiquantitative comparative analysis showed that 10-6 mol/L Cu2+ promotes the synthesis of Cry1Da and Cry1Ca (Additional file 1: Table S3). As previously discussed, Cu2+ caused ICP production to increase by 21 , and proteomic analysis revealed that the ICP are mainly composed of Cry1Ca, Cry1Ac, and Cry1Da. To investigate the effects of Cu2+ on ICP expression further, the transcripts of these three crystal proteins were analyzed by qRT-PCR. To determine whether or not changes in the proteome correlate with differences at the mRNA level, the transcripts of nine selected genes were analyzed by qRT-PCR, including five upregulated proteins (PhaR, BDH, EF-G, KAS II, and ALDH), three downregulated proteins (SHDA, ATPS, and PrkA), and three proteins showing no difference in proteomes [inosine-5-monophosphate dehydrogenase (IMPDH), small, acid-soluble spore protein B (SASPB), and ornithine aminotransferase (OAT)]. The growth curves obtained show that after 28 h of cultivation, higher bacterial concentrations were found in Cu than in CK. The pH variation curves show that PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/16989806 at 30?6 h of cultivation, the pH of the fermentation broth is higher in Cu than in CK (Fig. 5). Thus, we decided to extract the total RNA of the samples for qRT-PCR analysis at the time point of 31 h. Strain genome sequencing was limited to frame diagrams. The purpose of sequencing was performed to determine important functional genes than can help design gene-specific primer sequences. qRT-PCR results (Fig. 3) demonstrated that Cry1 Da and Cry1Ca are significantly upregulated at the transcriptional level, which confirms that appropriate amounts of Cu2+ have positive effect on 130 kDa Capecitabine ICP (Cry1Da and Cry1Ca) production. qRT-PCR showed positive correlations between changes at the translational and transcriptional levels of PhaR, BDH, EF-G, KAS II, ALDH, IMPDH, SASPB, and OAT but no correlation between changes at these levels of SHDA, ATPS, and PrkA.qRTPCR an.