Project description:On-demand biomanufacturing has the potential to improve healthcare and self- sufficiency during space missions. Cell-free transcription and translation reactions combined with DNA blueprints can produce promising therapeutics like bacteriophages and virus-like particles. However, how space conditions affect the synthesis and self-assembly of such complex multi- protein structures is unknown. Here, we characterize the cell-free production of infectious bacteriophage T7 virions under simulated microgravity. Rotation in a 2D-clinostat increased the number of infectious particles compared to static controls. Quantitative analyses by mass spectrometry, immuno-dot-blot and real-time PCR showed no significant differences in protein and DNA contents, suggesting enhanced self-assembly of T7 phages in simulated microgravity. While the effects of genuine space conditions on the cell-free synthesis and assembly of bacteriophages remain to be investigated, our findings support the vision of a cell-free synthesis-enabled “astropharmacy”.
Project description:We use Illumina sequencing to monitor mutations in the bacteriophage T7 genome in the presence of T7 DNA polymerase that has an altered exonuclease active site. These alterations include mutation of key residues in the exonuclease active site.
Project description:We have isolated and characterized several bacteriophages infecting Pseudomonas aeruginosa distantly related to Felix O1 virus and proposed they form a new subfamily named Felixounavirinae. The infectious cycle of bacteriophages belonging to this subfamily has not been studied yet in terms of gene expression. The present study reports the RNA-Seq analysis of bacteriophage PAK_P3 infecting PAK strain of P. aeruginosa.
Project description:We have isolated and characterized several bacteriophages infecting Pseudomonas aeruginosa distantly related to Felix O1 virus and proposed they form a new subfamily named Felixounavirinae. The infectious cycle of bacteriophages belonging to this subfamily has not been studied yet in terms of gene expression. The present study reports the RNA-Seq analysis of bacteriophage PAK_P4 infecting PAK strain of P. aeruginosa.
Project description:We have isolated and characterized several bacteriophages infecting Pseudomonas aeruginosa distantly related to Felix O1 virus and proposed they form a new subfamily named Felixounavirinae. The infectious cycle of bacteriophages belonging to this subfamily has not been studied yet in terms of gene expression. The present study reports the RNA-Seq analysis of bacteriophage PAK_P3 infecting PAK strain of P. aeruginosa. RNA profile of Host and Phage at 0min, 3.5min and 13 min after infection of Pseudomonas aeruginosa PAK strain with the Pseudomonas phage PAK P3. Three biological replicates for each time point.
Project description:Bacteriophage lambda is one of the most extensively studied organisms, and has been a primary model for understanding basic modes of genetic regulation. Here we examine the progress of lambda gene expression during phage development by ribosome profiling, and thereby provide a very high resolution view of lambda gene expression. The known genes are expressed in a predictable fashion, authenticating the analysis. But many previously unappreciated potential open reading frames become apparent in the expression analysis, revealing an unexpected complexity in the pattern of lambda gene function.
Project description:This SuperSeries is composed of the following subset Series: GSE16598: Tumor T1 Sectors GSE16599: Tumor T2 Sectors GSE16600: Tumor T3 Sectors GSE16601: Tumor T4 Sectors GSE16602: Tumor T5 Sectors GSE16603: Tumor T6 Sectors GSE16604: Tumor T7 Sectors GSE16605: Tumor T8 Sectors GSE16606: Tumor T9 Sectors GSE16607: Tumor T10 Sectors GSE16608: Tumor T11 Sectors GSE16609: Tumor T12 Sectors GSE16610: Tumor T13 Sectors GSE16611: Tumor T14 Sectors GSE16663: Tumor T15 Sectors GSE16664: Tumor T16 Sectors GSE16665: Tumor T17 Sectors GSE16667: Tumor T18 Sectors GSE16668: Tumor T19 Sectors GSE16669: Tumor T20 Sectors ROMA CGH experiments were conducted on four to six dissected sectors (S1-S6) from single primary ductal carcinomas (T1-T20). T1-T4 were analyzed by sectoring and analysis by ROMA. T5-T20 were analyzed using the Sector-Ploidy-Profiling (SPP) Approach. SPP involves macro-dissecting the tumor, flow-sorting nuclei by differences in total genomic DNA content and profiling the genome of the tumor subpopulations. The genomic DNA from each tumor was labeled with Cy5 and a reference fibroblast genome was labeled with Cy3 and cohybridized to a ROMA microarray (85K or 390K). Experiments were performed in color-reversal by dye-swapping samples and hybridizing the labeled DNA to a second microarray for T1-T15 experiments, but not for T16-T20 experiments.
