Project description:The promoter upstream from eryAI (eryAp) was cloned in front of EGFP in pIJ8660. This vector was integrated into the chromosome of K24-1, a clean host version of K41-135 (the overproducer strain obtained from Kosan Biosciences). This strain was then grown in shake flask culture in R5 medium. RNA was harvested up through three days. The 12 h RNA sample was used as the reference sample for all RNA hybridizations. A genomic DNA vs. 12 h control was also done to determine initial relative transcript expression.
Project description:The promoter upstream from eryAI (eryAp) was cloned in front of EGFP in pIJ8660. This vector was integrated into the chromosome of K24-1, a clean host version of K41-135 (the overproducer strain obtained from Kosan Biosciences). This strain was then grown in shake flask culture in R5 medium. RNA was harvested up through three days. The 12 h RNA sample was used as the reference sample for all RNA hybridizations. A genomic DNA vs. 12 h control was also done to determine initial relative transcript expression. Groups of assays that are related as part of a time series. Computed
Project description:The promoter upstream from eryAI (eryAp) was cloned in front of EGFP in pIJ8660. This vector was integrated into the chromosome of K24-1, a clean host version of K41-135 (the overproducer strain obtained from Kosan Biosciences). This strain was then grown in shake flask culture in R5 medium. RNA was harvested up through three days. The 12 h RNA sample was used as the reference sample for all RNA hybridizations. A genomic DNA vs. 12 h control was also done to determine initial relative transcript expression. Groups of assays that are related as part of a time series. Keywords: time_series_design
Project description:The present work aimed at providing noval reference genes (RGs) for streptomycetes by global quantitative analysis of gene expression profile. By using the time-series microarray data obtained in different culture medium (SMM in the present work and modified R5 from publication), the stably expressed genes of S. coelicolor were screened. Further statistical, bioinformatic and biological function analysis picked out 13 candidate RGs. According to qRT-PCR assays, 5 genes with high stability were selected and used for validation in other streptomycetes to assess their prevalence. Additionally, the absolute gene expression level reflcted by RNA-seq was also taken into consideration in the present work to guide the appropriate RGs selection for streptomycetes.
