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Transgenic yeast (pAG305GPD-AAE13) vs. control yeast (pAG305GPD-ccdB)

ABSTRACT: The yeast expression vectors used to express AAE13 was constructed as follows: AAE13 (At3g16170) was amplified from the vector pENTR-AAE13. The amplified product was cloned into pCR®8⁄GW⁄TOPO® (Invitrogen). The resulting plasmid was combined with the Advanced Gateway destination vectors pAG305GPD-ccdB (Addgene, Boston, MA) in an attL X attR recombination reaction to generate the vectors pAG305GPD-AAE13. The integrating vector pAG305GPD-AAE13 was introduced into WAT11, which was also transformed empty vector pAG305GPD-ccdB as control (B1, B2, B3 and B4). The transgenic and control yeast cells were cultured in SD-drop out medium at 30 °C under shaken conditions, malonate was fed into cells at final concentration 0.2 mM when the value of OD600 reached at 0.1. The cells were harvested at mid-log phase.

ORGANISM(S): Saccharomyces cerevisiae

PROVIDER: GSE52701 | GEO | 2013/11/26

SECONDARY ACCESSION(S): PRJNA229864

REPOSITORIES: GEO

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Transgenic yeast (pAG305GPD-AAE13) vs. control yeast (pAG305GPD-ccdB)

Project description:The yeast expression vectors used to express AAE13 was constructed as follows: AAE13 (At3g16170) was amplified from the vector pENTR-AAE13. The amplified product was cloned into pCRM-BM-.8M-bM-^AM-^DGWM-bM-^AM-^DTOPOM-BM-. (Invitrogen). The resulting plasmid was combined with the Advanced Gateway destination vectors pAG305GPD-ccdB (Addgene, Boston, MA) in an attL X attR recombination reaction to generate the vectors pAG305GPD-AAE13. The integrating vector pAG305GPD-AAE13 was introduced into WAT11, which was also transformed empty vector pAG305GPD-ccdB as control (B1, B2, B3 and B4). The transgenic and control yeast cells were cultured in SD-drop out medium at 30 M-BM-0C under shaken conditions, malonate was fed into cells at final concentration 0.2 mM when the value of OD600 reached at 0.1. The cells were harvested at mid-log phase. Two-condition experiment, Transgenic yeast (pAG305GPD-AAE13) vs. control yeast (pAG305GPD-ccdB). Biological replicates: 4 control replicates, 4 transgenic replicates.

2013-11-26 | E-GEOD-52701 | biostudies-arrayexpress

NCI-H1299 cell line transfected with human DENND2D and vector

Project description:Gene expression for NCI-H1299 cell line transfected with human DENND2D and vector (pcDNA3.1/V5-His TOPO TA vector) respectively.

2010-05-04 | GSE16890 | GEO

Expression data from 26972c yeast ± bHLHm1 (SAT1)

Project description:26972c yeast cells were transformed with either empty vector (pYES3) or pYES3:Gm:bHLHm1. Cells were grown on low ammonium concentrations to observe transcriptional changes in the yeast genome in response to the soybean bHLHm1 transcription factor.

2014-03-12 | GSE55804 | GEO

Transcriptomic response of the yeast Saccharomyces cerevisiae EC1118® during wine alcoholic fermentation under unusual nutrient starvations.

Project description:In order to asses yeast EC1118® strain expression changes during wine alcoholic fermentation triggered by various nutrient starvations, this experiment describes the gene expression under micronutrient starvations that lead to yeast cell death (oleic acid starvation, ergosterol starvation, pantothenic acid starvation and nicotinic starvation) or allow the maintenance of yeast viability (nitrogen starvation).

2017-07-30 | GSE95152 | GEO

Yeast genomic expression patterns in response to zymolyase treatment

Project description:The evaluation of zymolyase treatment using yeast gene expression comparison analysis. The yeast BY4743 strain was used for this study. Yeast cells were treated with Zymolyase (final conc. 300 U/ml), and incubated at 37 degree for ten minutes. Digestion process was not provided for control sample. Total RNA was isolated with commercial kit (FastRNA Pro Red kit, Q-Biogene), and amplified RNA (aRNA) was synthesized with 3'IVT Express kit (Affymetrix). All samples were hybridized with Gene Chip (Yeast Genome 2.0 Array, Affymetrix), then each array chip was scanned by GeneChip Sanner 3000 (Affymetrix).

2013-03-21 | GSE36955 | GEO

Yeast genomic expression patterns in response to type B trichothecenes

Project description:The evaluation of mycotoxicity of type B trichothecenes using a yeast gene expression comparison analysis. The yeast BY4743 derivative PDR5 mutant was used for this study. The yeast cells were treated with trichothecene mycotoxins, and incubated at 25 degree for two hours, respectively. Total RNA was isolated with commercial kit (FastRNA Pro Red kit, Q-Biogene), and amplified RNA (aRNA) was synthesized with 3'IVT Express kit (Affymetrix). All samples were hybridized with Gene Chip (Yeast Genome 2.0 Array, Affymetrix), then each array chip was scanned by GeneChip Sanner 3000 (Affymetrix).

2012-09-12 | GSE36954 | GEO

NCI-H1299 cell line transfected with human DENND2D and vector

Project description:Gene expression for NCI-H1299 cell line transfected with human DENND2D and vector (pcDNA3.1/V5-His TOPO TA vector) respectively. The microarray experiment was designed to perform four replicates for each H1299-DENND2D and H1299-vector sample. cRNA used in replication 1 and 2 was from the same label reaction to perform the hybridization replicates.

2010-05-27 | E-GEOD-16890 | biostudies-arrayexpress

Comparison of genome wide binding of S. cerevisiae condensin, topo II and cohesin by ChIP

Project description:The chromosomal condensin complex gives metaphase chromosomes structural stability. In addition, condensin is required for sister chromatid resolution during their segregation in anaphase. How condensin promotes chromosome resolution is poorly understood. Chromosome segregation during anaphase also fails after inactivation of topoisomerase II (topo II), the enzyme that removes catenation between sister chromatids left behind after completion of DNA replication. This has led to the proposal that condensin promotes DNA decatenation, but direct evidence for this is missing and alternative roles for condensin in chromosome resolution have been suggested. Using the budding yeast rDNA as a model, we now show that anaphase bridges in a condensin mutant are resolved by ectopic expression of a foreign (Chlorella virus) but not endogenous topo II. This suggests that catenation prevents sister rDNA segregation, and that yeast topo II is ineffective in decatenating the locus without condensin. Condensin and topo II colocalize along both rDNA and euchromatin, consistent with coordination of their activities. We investigate the physiological consequences of condensin-dependent rDNA decatenation and find that late decatenation determines the late segregation timing of this locus during anaphase. Regulation of decatenation therefore provides a means to finetune segregation timing of chromosomes in mitosis. Keywords: ChIP-chip, Cell Cycle

2008-07-18 | GSE11988 | GEO

QUILT-2.025 NANT Neoepitope Yeast Vaccine (YE-NEO-001): Adjuvant Immunotherapy Using a Personalized Neoepitope Yeast-Based Vaccine To Induce T-Cell Responses In Subjects W/ Previously Treated Cancers.

Project description:This is a phase 1 study to evaluate the safety, the Recommended Phase 2 Dose (RP2D), and preliminary efficacy of a personalized neoepitope yeast-based vaccine, YE-NEO-001, in subjects who have completed potentially curative therapy for their solid cancer and who would otherwise be entering a period of surveillance for recurrent disease.

| 2275044 | ecrin-mdr-crc

Myometrial cells expressing CREB, CREM alpha, CREM tau2alpha, ATF2 or the ATF2-small gene

Project description:Human myometrial cells taken from biopsies of pre-menopausal non-cancerous non-pregnant uteri were cultured in complete MEM D-valine medium plus 10% FCS, and stably-transfected with the pcDNA3.1/V5-His TOPO vector (Invitrogen) harbouring either CREB, CREM alpha, CREM tau2alpha, ATF2, ATF2-small gene or a control empty-vector. Total RNA was extracted using TRI-reagent (Sigma) and RNeasy columns (Qiagen), and 10 ug used in target preparation for array hybridization. Keywords: other

2004-02-21 | GSE1059 | GEO

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