Project description:100mM EMS was added to mid-log phase Halobacterium NRC-1 cultures. After constant stress of EMS for 30 minutes, cultures were spun down and pellets were re-suspended in same volume of GN101 media. Cultures were then harvested by centrifugation and pellets were snap frozen on a dry-ice ethanol bath. Samples for RNA preparation were collected during recovery time points at 0, 10, 20, 30, 40, 60 and 120 minutes. Keywords: stress response, dose response
Project description:We performed DNA Adenine Methyltransferase Identification (DamID) to map the contacts between the genome and the nuclear lamina in the EMS lineage of early embryos. Dam::EMR-1 and GFP::Dam fusion proteins were expressed from single-copy transgenes inserted into an intergenic region on chromosome II. To induce a short burst of transcription of the transgenes, we used the med-1 promoter, which is active in the EMS cell of 4 cell-stage embryos and repressed in the descendants of EMS.
Project description:Granulosa cells are essential to the growth, development, and maturation of oocytes and follicles. In patients with EMs-related infertility, follicle maturation disorders, poor oocyte quality may be closely related to GCs in the follicle. However, the molecular biological mechanism of GCs in EMs-related infertility has not been reported. Therefore, we tried to explore the possible causes of EMs-related infertility through the study of ovarian GCs in patients with EMs-related infertility and tubal factor infertility.
Project description:100mM EMS was added to mid-log phase Halobacterium NRC-1 cultures. After constant stress of EMS for 30 minutes, cultures were spun down and pellets were re-suspended in same volume of GN101 media. Cultures were then harvested by centrifugation and pellets were snap frozen on a dry-ice ethanol bath. Samples for RNA preparation were collected during recovery time points at 0, 10, 20, 30, 40, 60 and 120 minutes. 16 samples (8 samples from EMS perturbed and 8 non-perturbed controls) were analyzed on replicate arrays (dye-flips) all against the same standard reference sample.
Project description:This study employed an unbiased forward genetic screen to identify suppressors of the maternal-effect lethal phenotype caused by tost-1 dysfunction in Caenorhabditis elegans. The temperature-sensitive tost-1(xf196 ts) mutant exhibits complete embryonic lethality at the restrictive temperature (25°C), providing a robust screening platform. Following EMS mutagenesis of L4 larvae, we screened F2 progeny for viable individuals at 25°C, indicating suppression of the lethal phenotype. After backcrossing to confirm heritability, we performed whole genome sequencing on 21 suppressor strains to identify causative mutations. Additionally we also sequenced the strain used for the mutagenesis as the reference genome.