Morphological differentiation and secondary metabolism responses of Streptomyces coelicolor A3(2) to simulated microgravity based on comparative transcriptomics.
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ABSTRACT: In order to gain a better understanding of the effects of Streptomyces coelicolor A3(2) wild type strain exposed to simulated microgravity condition by clinorotation, we constructed a whole genome microarray to screen simulated microgravity sensitive genes using the Agilent eArray 5.0 program. The genechip was custom designed according to the manufacturer’s recommendations (Agilent, 8*15K). The genome sequences were downloaded from: http://www.ncbi.nlm.nih.gov/genome?Db =genome&Cmd = Search&Term = NC_003888, NC_003903.1, NC_003904.1. Each gene was represented by one 60-nt oligonucleotide probe, and 680 genes associated with morphological differentiation and secondary metabolism out of the total 8116 genes were replicated 9 times each.
Project description:In order to gain a better understanding of the effects of Streptomyces coelicolor A3(2) wild type strain exposed to simulated microgravity condition by clinorotation, we constructed a whole genome microarray to screen simulated microgravity sensitive genes using the Agilent eArray 5.0 program. The genechip was custom designed according to the manufacturer’s recommendations (Agilent, 8*15K). The genome sequences were downloaded from: http://www.ncbi.nlm.nih.gov/genome?Db =genome&Cmd = Search&Term = NC_003888, NC_003903.1, NC_003904.1. Each gene was represented by one 60-nt oligonucleotide probe, and 680 genes associated with morphological differentiation and secondary metabolism out of the total 8116 genes were replicated 9 times each.
Project description:In order to gain a better understanding of the effects of Streptomyces coelicolor A3(2) wild type strain exposed to spaceflight condition, we constructed a whole genome microarray to screen simulated microgravity sensitive genes using the Agilent eArray 5.0 program. The genechip was custom designed according to the manufacturer’s recommendations (Agilent, 8*15K). The genome sequences were downloaded from: http://www.ncbi.nlm.nih.gov/genome?Db =genome&Cmd = Search&Term = NC_003888, NC_003903.1, NC_003904.1. Each gene was represented by one 60-nt oligonucleotide probe, and 680 genes associated with morphological differentiation and secondary metabolism out of the total 8116 genes were replicated 9 times each.
Project description:In order to gain a better understanding of the effects of Streptomyces coelicolor A3(2) wild type strain exposed to simulated microgravity condition by clinorotation, we constructed a whole genome microarray to screen simulated microgravity sensitive genes using the Agilent eArray 5.0 program. The genechip was custom designed according to the manufacturer’s recommendations (Agilent, 8*15K). The genome sequences were downloaded from: http://www.ncbi.nlm.nih.gov/genome?Db =genome&Cmd = Search&Term = NC_003888, NC_003903.1, NC_003904.1. Each gene was represented by one 60-nt oligonucleotide probe, and 680 genes associated with morphological differentiation and secondary metabolism out of the total 8116 genes were replicated 9 times each. Cells of Streptomyces coelicolor A3(2) wild type strain were harvested from the colonies grown in JCM42 agar medium at 28°C for 144h (6d) under simulated microgravity (by a rotating clinostat) and 1g control conditions respectively. Total RNA was extracted to conduct the microarray hybridization experiments for two color (Cy3/Cy5).
Project description:In order to gain a better understanding of the effects of Streptomyces coelicolor A3(2) wild type strain exposed to simulated microgravity condition by clinorotation, we constructed a whole genome microarray to screen simulated microgravity sensitive genes using the Agilent eArray 5.0 program. The genechip was custom designed according to the manufacturerM-bM-^@M-^Ys recommendations (Agilent, 8*15K). The genome sequences were downloaded from: http://www.ncbi.nlm.nih.gov/genome?Db =genome&Cmd = Search&Term = NC_003888, NC_003903.1, NC_003904.1. Each gene was represented by one 60-nt oligonucleotide probe, and 680 genes associated with morphological differentiation and secondary metabolism out of the total 8116 genes were replicated 9 times each. Cells of Streptomyces coelicolor A3(2) wild type strain were harvested from the colonies grown in JCM42 agar medium at 28M-BM-0C for 96h (4d) under simulated microgravity and 1g control conditions. Total RNA was extracted to conduct the microarray hybridization experiments. Three independent experiments were performed in which ASM_1A, ASM_2A, ASM_3A were the test group samples and ANG_1A, ANG_2A, ANG_3A were the control group samples. Differentially expressed genes were selected with PM-oM-<M-^\0.05, FCM-bM-^IM-%2.0 by T-test methods using the independent three sets data.
Project description:In order to gain a better understanding of the effects of Streptomyces coelicolor A3(2) wild type strain exposed to spaceflight condition, we constructed a whole genome microarray to screen simulated microgravity sensitive genes using the Agilent eArray 5.0 program. The genechip was custom designed according to the manufacturerM-bM-^@M-^Ys recommendations (Agilent, 8*15K). The genome sequences were downloaded from: http://www.ncbi.nlm.nih.gov/genome?Db =genome&Cmd = Search&Term = NC_003888, NC_003903.1, NC_003904.1. Each gene was represented by one 60-nt oligonucleotide probe, and 680 genes associated with morphological differentiation and secondary metabolism out of the total 8116 genes were replicated 9 times each. Cells of Streptomyces coelicolor A3(2) wild type strain were harvested from the colonies grown in JCM42 agar medium at 23M-BM-10.5M-BM-0C for 16.5 days in spaceflight experiments. Total RNA was extracted to conduct the microarray experiments. The samples labeled in SP12_1 were from the M-bM-^@M-^\M-NM-<g-positionM-bM-^@M-^] and C07_1 from the M-bM-^@M-^\simulated 1g-positionM-bM-^@M-^] inside the BIOBOX for spaceflight during Shenzhou-8 space mission. The samples labeled in DB3_1 were from static 1g position on gound as the control. SP12_1 was test sample; C07_1 and DB3_1 were both samples for control.
Project description:In order to gain a better understanding of the mechanism of increased cell death triggered by deficient GPI anchoring, we constructed a whole genome microarray using the Agilent eArray 5.0 program.The chip specification is 8×15K. The genome sequences were downloaded from the following site:http://www.ncbi.nlm.nih.gov/genome?Db=genome&Cmd=Search&Term=txid330879[orgn]. Each gene was represented by one 60-nt oligonucleotide probe, and 358 genes out of the 9630 total were replicated 14 times.Using this microarray,we got 3274 genes which were induced or reduced at least 1.5 fold in the afpig-a mutant comparing to the wild-type.
Project description:In order to gain a better understanding of the mechanism of increased cell death triggered by deficient GPI anchoring, we constructed a whole genome microarray using the Agilent eArray 5.0 program.The chip specification is 8M-CM-^W15K. The genome sequences were downloaded from the following site:http://www.ncbi.nlm.nih.gov/genome?Db=genome&Cmd=Search&Term=txid330879[orgn]. Each gene was represented by one 60-nt oligonucleotide probe, and 358 genes out of the 9630 total were replicated 14 times.Using this microarray,we got 3274 genes which were induced or reduced at least 1.5 fold in the afpig-a mutant comparing to the wild-type. Mycelia of the afpig-a mutant and the wild-type were harvested from strains grown in liquid CM at 37M-BM-0C for 36h with shaking at 200 rpm and ground by hand.Total RNA was extracted to do the hybridization.cRNAs were labled with Cy3 NHS ester.afpig-a_37 was sample, afpig_37_1a,afpig_37_2a,afpig_37_3a and afpig_37_4a were biological repeats. w_37 was control,w_37_1a,w_37_2a,w_37_3a and w_37_4a were biological repeats.Four independent experiments were performed. Each time we got a set of differentilly expressed genes.Differentially expressed genes were selected with PM-bM-^IM-$0.05, FCM-bM-^IM-%1.5 by T-test methods using the independent four sets data.
Project description:Cellular and molecular dynamics of human cells are constantly affected by gravity. Alteration of the gravitational force disturbs the cellular equilibrium, which might modify physiological and molecular characteristics. Nevertheless, biological responses of cancer cells to reduced gravitational force remains obscure. Here, we aimed to comprehend not only transcriptomic patterns but drug responses of colorectal cancer (CRC) under simulated microgravity. We established four organoids directly from CRC patients, and organoids cultured in 3D clinostat were subjected to genome wide expression profiling and drug library screening. Our observations revealed changes in cell morphology and an increase in cell viability under simulated microgravity compared to their static controls. Transcriptomic analysis highlighted a significant dysregulation in the TBC1D3 family of genes. The upregulation of cell proliferation observed under simulated microgravity conditions was further supported by enriched cell cycle processes, as evidenced by the functional clustering of mRNA expressions using cancer hallmark and gene ontology terms. Our drug screening results indicated an enhanced response rate to 5-FU under conditions of simulated microgravity, suggesting potential implications for cancer treatment strategies in simulated microgravity.
Project description:To investigate the differential action between resistance and susceptible cultivars, we examined genome wide expression levels at five time points after Xag-inoculation using microarray. The soybean microarray was designed using the server-based eArray platform (http://earray.chem.agilent.com/earray/) from Agilent Technologies (Wolber et al., 2006). The current slide layout consists of four arrays of >44,000 features, including a set of Agilent's positive and negative control features. An oligonucleotide microarray containing 42,564 unique probes (60-mer) was created from 33,574 unigenes from Glycine max database (http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?id=3847) (May 2009) as templates for probe design.
Project description:In space, multiple unique environmental factors, particularly microgravity and space radiation, pose constant threat to the astronaut’s health. To gain insight into the role of miRNAs and lncRNAs in response to radiation and microgravity, we analyzed RNA expression profiles in human lymphoblastoid TK6 cells incubated for 24 h in static condition or in rotating condition to stimulate microgravity in space after 2 Gy γ-ray irradiation. Expression of 14 lncRNAs and 17 mRNAs was found to be significantly down-regulated in the simulated microgravity condition. In contrast, irradiation up-regulated the expression of 55 lncRNAs and 56 mRNAs, while only one lncRNA, but no mRNA, was down-regulated. Furthermore, 2 miRNAs, 70 lncRNAs, and 87 mRNAs showed significantly altered expression under simulated microgravity after irradiation, and these changes were independently induced by irradiation and simulated microgravity. Together, our results indicate that simulated microgravity and irradiation additively and independently alter the expression of RNAs and their target genes in human lymphoblastoid cells.