An electrophoretic mobility shift assay can be used to study the binding of proteins to a segment of DNA. In the results shown here, an EMSA was used to examine the requirements for the binding of RNA polymerase |l (from eukaryotic cells) to the promoter of a protein-encoding gene. The assembly of general transcription factors and RNA polymerase Il at the core promoter is described in Week 4. In this experiment, the segment of DNA containing a promoter sequence was 1100 bp in length. The fragment was mixed with various combinations of proteins and then subjected to an EMSA. Lane 1: No proteins added Lane 2: TFIID Lane 3: TFIIB Lane 4: RNA polymerase Il Lane 5: TFIID + TFIIB Lane 6: TFIID + RNA polymerase II 2 3 4 7 Lane 7: TFIID + TFIIB + RNA polymerase II 1100 bp Explain the results.

An electrophoretic mobility shift assay can be used to study the binding of proteins to a segment of DNA. In the results shown here, an EMSA was used to examine the requirements for the binding of RNA polymerase |l (from eukaryotic cells) to the promoter of a protein-encoding gene. The assembly of general transcription factors and RNA polymerase Il at the core promoter is described in Week 4. In this experiment, the segment of DNA containing a promoter sequence was 1100 bp in length. The fragment was mixed with various combinations of proteins and then subjected to an EMSA. Lane 1: No proteins added Lane 2: TFIID Lane 3: TFIIB Lane 4: RNA polymerase Il Lane 5: TFIID + TFIIB Lane 6: TFIID + RNA polymerase II 2 3 4 7 Lane 7: TFIID + TFIIB + RNA polymerase II 1100 bp Explain the results.

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Description: An electrophoretic mobility shift assay can be used tostudy the binding of proteins to a segment of DNA. Inthe results shown here, an EMSA was used toexamine the requirements for the binding of RNApolymerase |l (from eukaryotic cells) to the promote

Human Heredity: Principles and Issues (MindTap Course List)

11th Edition

ISBN:9781305251052

Author:Michael Cummings

Publisher:Michael Cummings

Chapter15: Genomes And Genomics

Section: Chapter Questions

Problem 13QP

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Certain hormones, such as epinephrine, can increase the levels ofcAMP within cells. Let’s suppose you pretreat cells with or withoutepinephrine and then prepare a cell extract that contains theCREB protein.You then use an electrophoretic mobility shift assay to analyzethe ability of the CREB protein to bind to a DNA fragmentcontaining a cAMP response element (CRE). Describe what theexpected results would be.
explain please: Spike-Driven Syncytia Formation Is Coupled to S20 Fragment Generation in the Presence of ACE2. To investigate the functional and biochemical signatures of spike (S) protein upon engaging its receptor ACE2 in live cells, we first utilized a cell–cell fusion assay to obtain potentially cleaved spike protein products from syncytia . In this system, HEK293T cells transfected with plasmids encoding WT spike readily formed syncytia after coculturing with HEK293T cells expressing human and Vero E6-ACE2, as well as colorectal adenocarcinoma Caco-2 and lung adenocarcinoma Calu-3 cells expressing endogenous ACE2. We then collected these adherent syncytia and immunoblotted for cleaved spike species using a rabbit polyclonal antibody specifically detecting the S2 ectodomain through standard reducing Tris-glycine sodium dodecyl sulfate polyacrylamide gel electrophoresis. This experiment revealed that spike expression in HEK293T cells displayed full-length S and autocleaved S2…
E30. An electrophoretic mobility shift assay can be used to study the binding of proteins to a segment of DNA. In the experiment shown here, an EMSA was used to examine the requirements for the bind- ing of RNA polymerase II (from eukaryotic cells) to the promoter of a protein-encoding gene. The assembly of general transcription factors and RNA polymerase II at the core promoter is described in Chapter 12 (Figure 12.14). In this experiment, the segment of DNA containing a promoter sequence was 1100 bp in length. The fragment was mixed with various combinations of proteins and then subjected to an EMSA. Lane 1: No proteins added Lane 2: TFID Lane 3: TFIIB Lane 4: RNA polymerase I| Lane 5: TFID + TFIIB Lane 6: TFID + RNA 1 2 4 5 6 polymerase II| Lane 7: TFIID + TFIIB + RNA polymerase I| 1100 bp Explain which proteins (TFIID, TFIIB, or RNA polymerase II) are able to bind to this DNA fragment by themselves. Which transcrip- tion factors (i.e., TFIID or TFIIB) are needed for the binding of…
Another mutant for of the Lac promoter demonstrates the characteristics below compared to the wild-type promoter. The RNA Polymerase Holoenzyme binds tighter to the mutant promoter, but has reduced promoter activity in an assay with the promoter fuse to the gene for Green Fluorescent Protein (GFP). Provide a plausible explanation for this data. Lac Promoter Activity punog Binding of RNA Pol Holoenzymes to Promoters 120 100 80 60 40 20 0 0 20 40 Lac Promoter X 60 [DNA] nM 80 100 120 GFP Fluorecence 100 90 80 70 60 50 40 30 20 10 0 Lac Promoter X
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