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- How did the following in-vitro transcription assay utilizing the rrnB promoter help elucidate the UP story? Explain the experiment, the controls [lacUV5, RNA-1], the various DNA promoters, and the data [including any artifact]. (a) Copyright © The McGraw-H Comperies, Inc. Permission required for reproduction or display (b) (c) WT Was-type RNAP 0-235 lac 88 SUB-41 UVS vector la -88 SUB41 UVS vector lacUV5- mm P1 RNA-1 Gel A..... 12345678910 12345678910 Roset, Aindecagnition sla gel B. P20G WT P205C WTR265C -88 SUB-SUB US 123456 7491011121314 lacuVS ma Pt- PINA-1 prostars DNA binding by the alpha subunit of FA polymerase Now 1990) 1. 2. p. 1408 0 AAAS

- How did the following in-vitro transcription assay utilizing the rrnB promoter help elucidate the UP story? Explain the experiment, the controls [lacUV5, RNA-1], the various DNA promoters, and the data [including any artifact]. (a) Copyright © The McGraw-H Comperies, Inc. Permission required for reproduction or display (b) (c) WT Was-type RNAP 0-235 lac 88 SUB-41 UVS vector la -88 SUB41 UVS vector lacUV5- mm P1 RNA-1 Gel A..... 12345678910 12345678910 Roset, Aindecagnition sla gel B. P20G WT P205C WTR265C -88 SUB-SUB US 123456 7491011121314 lacuVS ma Pt- PINA-1 prostars DNA binding by the alpha subunit of FA polymerase Now 1990) 1. 2. p. 1408 0 AAAS

Human Anatomy & Physiology (11th Edition)
Human Anatomy & Physiology (11th Edition)
11th Edition
ISBN:9780134580999
Author:Elaine N. Marieb, Katja N. Hoehn
Publisher:Elaine N. Marieb, Katja N. Hoehn
Chapter1: The Human Body: An Orientation
Section: Chapter Questions
Problem 1RQ: The correct sequence of levels forming the structural hierarchy is A. (a) organ, organ system,...
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Molecular biology, please explain in detail
### In-Vitro Transcription Assay Utilizing the rrnB Promoter to Elucidate the UP Story How did the following in-vitro transcription assay utilizing the rrnB promoter help elucidate the UP story? Explain the experiment, the controls [lacUV5, RNA-1], the various DNA promoters, and the data [including any artifact]. #### Experiment Details and Explanation: In-vitro transcription assays are conducted to study how various elements of a promoter regulate transcription. Here, the rrnB promoter was used to understand the UP element's influence on transcription efficiency. Key components assessed include RNA polymerase (RNAP) activity, the roles of different promoters, and control DNA sequences. #### Controls: 1. **lacUV5**: Serves as a baseline promoter to compare transcriptional activity. 2. **RNA-1**: Another control used to ensure the specificity and efficiency of the transcription reaction. The data is depicted using three autoradiographs (labeled as a, b, and c), which represent the results of in-vitro transcription assays under varying conditions and with different DNA templates. #### Gel Analysis: **Figure (a)**: - **Gal Sub and LacUV5 Vector**: This gel shows transcription activity with different DNA templates and RNAP. - There are strong, visible bands for lacUV5 and rrnB suggesting active transcription. - Substantial activity for SUB and -88, indicating the influence of upstream and core promoter regions. **Figure (b)**: - **q282 Wild-Type and LacUV5 Vector**: Similar setup as Figure (a) but using a different wild-type promoter. - The band intensities and their positions help infer the activity levels of q282 relative to lacUV5. - Strong transcriptional activity observed, particularly for lacUV5. **Figure (c)**: - **WT, R285G, and R285C Mutants with LacUV5**: Analysis using wild-type and mutant promoters to hypothesize the influence of specific mutations on transcription. - Comparison between WT, R285G, and R285C displays varying degrees of transcription efficiency. - LacUV5 again shows high activity, while mutant promoters (e.g., -88 and SUB) illustrate the promoter strength differences influenced by mutations. #### Conclusion: This in-vitro transcription assay helped elucidate the role of the UP element in enhancing transcription efficiency by comparing various promoters and mutant forms. The use of controls
Transcribed Image Text:### In-Vitro Transcription Assay Utilizing the rrnB Promoter to Elucidate the UP Story How did the following in-vitro transcription assay utilizing the rrnB promoter help elucidate the UP story? Explain the experiment, the controls [lacUV5, RNA-1], the various DNA promoters, and the data [including any artifact]. #### Experiment Details and Explanation: In-vitro transcription assays are conducted to study how various elements of a promoter regulate transcription. Here, the rrnB promoter was used to understand the UP element's influence on transcription efficiency. Key components assessed include RNA polymerase (RNAP) activity, the roles of different promoters, and control DNA sequences. #### Controls: 1. **lacUV5**: Serves as a baseline promoter to compare transcriptional activity. 2. **RNA-1**: Another control used to ensure the specificity and efficiency of the transcription reaction. The data is depicted using three autoradiographs (labeled as a, b, and c), which represent the results of in-vitro transcription assays under varying conditions and with different DNA templates. #### Gel Analysis: **Figure (a)**: - **Gal Sub and LacUV5 Vector**: This gel shows transcription activity with different DNA templates and RNAP. - There are strong, visible bands for lacUV5 and rrnB suggesting active transcription. - Substantial activity for SUB and -88, indicating the influence of upstream and core promoter regions. **Figure (b)**: - **q282 Wild-Type and LacUV5 Vector**: Similar setup as Figure (a) but using a different wild-type promoter. - The band intensities and their positions help infer the activity levels of q282 relative to lacUV5. - Strong transcriptional activity observed, particularly for lacUV5. **Figure (c)**: - **WT, R285G, and R285C Mutants with LacUV5**: Analysis using wild-type and mutant promoters to hypothesize the influence of specific mutations on transcription. - Comparison between WT, R285G, and R285C displays varying degrees of transcription efficiency. - LacUV5 again shows high activity, while mutant promoters (e.g., -88 and SUB) illustrate the promoter strength differences influenced by mutations. #### Conclusion: This in-vitro transcription assay helped elucidate the role of the UP element in enhancing transcription efficiency by comparing various promoters and mutant forms. The use of controls
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