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Comparison of Transcription Profiles of M. smegmatis overexpressing wild type and phopshorylation defective PknK

ABSTRACT: We have shown that elevated levels of wild-type M. tuberculosis PknK (PknK Mtb), but not phosphorylation-defective PknKMtb, in Mycobacterium smegmatis cause significant retardation of growth rate and altered colony morphology (Malhotra et al., 2012). LIX79 (WT PknK) and LIX80 (PknK K55M mutant) M. smegmatis strains will be grown in LB and induced with 0.2% acetamide for the induction of wild type or phosphorylation defective pknK gene. RNA and Protein will be isolated from cultures pelleted at times—24 h (24h) and 96 h (96 h) post induction. M. smegmatis strain LIX70 containing the empty vector will be induced at the same time-points and used as a control in hybridization. Two independent replicate experiments will be done.

ORGANISM(S): Mycolicibacterium smegmatis

PROVIDER: GSE180348 | GEO | 2022/03/10

REPOSITORIES: GEO

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Project description:Total transcriptome analyses were carried out to investigate changes in transcript patterns in the Mycobacterium smegmatis wild type strain SMR5 due to nitrogen starvation and to expand the knowledge about the role of the two transcriptional regulators of nitrogen metabolism, namely GlnR and AmtR, in these processes. A first experiment revealed enhanced transcript levels of 284 genes and reduced transcripts of 231 genes in the wild type under nitrogen starvation compared to nitrogen surplus. When glnR deletion strain MH1 was compared to the wild type under nitrogen starvation, decreased transcript levels of 125 genes were detected, indicating that these are activated by GlnR due to nitrogen limitation. Comparing amtR deletion strain YL1 to the wild type under nitrogen starvation, enhanced transcript levels of 2 genes were found, indicating that they are repressed by AmtR under nitrogen surplus. A comparison of YL1 and wild type under surplus, as well as a comparison of YL1 under nitrogen surplus and starvation and of MH1 under nitrogen surplus and starvation were carried out as additional control experiments. It can be concluded that GlnR is the master regulator of nitrogen control in M. smegmatis and that AmtR fulfills only a small, subordinate role in the regulation of an operon. This SuperSeries is composed of the following subset Series: GSE30033: Mycobacterium smegmatis - Comparison of glnR deletion strain MH1 under nitrogen surplus and starvation GSE30231: Mycobacterium smegmatis - Comparison of wild type SMR5 and amtR deletion strain YL1 under nitrogen starvation GSE30232: Mycobacterium smegmatis - Comparison of amtR deletion strain YL1 under nitrogen surplus and starvation GSE30233: Mycobacterium smegmatis - Comparison of wild type SMR5 under nitrogen surplus and starvation GSE30234: Mycobacterium smegmatis - Comparison of wild type SMR5 and glnR deletion strain MH1 under nitrogen starvation GSE30235: Mycobacterium smegmatis - Comparison of wild type SMR5 and amtR deletion strain YL1 under nitrogen surplus Refer to individual Series

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Comparison of Transcription Profiles of M. smegmatis overexpressing wild type and phopshorylation defective PknK

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