Strength 22. You are analyzing a new human neuronal enhancer. You cloned the enhancer in front of a minimal promoter, transfected a neuronal cell line, and quantified GFP expression. You then performed the deletion analysis shown on the right. Which letter designates the minimal region where enhancer activity is located? min. Novel enhancer prom. GFP of expression +++ +/- +++ +/- +++ a. a b. b C. C +++ +/- d. d e. both b and c are needed

Strength 22. You are analyzing a new human neuronal enhancer. You cloned the enhancer in front of a minimal promoter, transfected a neuronal cell line, and quantified GFP expression. You then performed the deletion analysis shown on the right. Which letter designates the minimal region where enhancer activity is located? min. Novel enhancer prom. GFP of expression +++ +/- +++ +/- +++ a. a b. b C. C +++ +/- d. d e. both b and c are needed

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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1.Your goal is to design a strategy to create thisfinal pDHFR plasmid for fusion protein expressionfrom the materials available:You have an empty pET21a expression vector and another vectorthat contains the fusion protein nucleotide sequence (pBluescript + GST-DHFR-His). (#) indicatesbp location ofenzyme cut: a.EcoRIonly, HindIII only, NotIonly, EcoRI and HindIII, EcoRI and NotI, or HindIII and NotI? b.Briefly explain the rationale for your selection. Please connect your rationale to the need for compatible ends without any further manipulation and required directionality of the DHFR fusion protein coding sequence relative to the T7 promoter that will drive its expression. c.What is the final size(kb)of your desired pDHFR plasmid ligation product based onyour design? d. Complete the chart providedto indicate thesize of all fragments(in bp)resultingfrom complete digestion of each plasmid givenyour strategy, andthe fragment(in kb)from each you would isolate for ligation.Please mind…
Time (h) 0 1 3 6 12 Mouse thymus cells were treated with 10 micromolar dexamethasone to induce apoptosis. At the indicated times the chromosomal DNA was extracted and analyzed by gel electrophoresis, which separates the DNA fragments based upon size differences. When cells undergo apoptosis their DNA is cut into fragments that are approximately multiples of 200 base pairs. This results in a characteristic “ladder" appearance when the fragments are separated by gel electrophoresis, as shown above. Based upon what you know about eukaryotic chromatin, explain why this pattern of DNA fragments is obtained. 6.
Please answer fast If the plasmid Lac operon has a mutated lacO operator gene that prevents the repressor from binding, which of the following will occur when lactose is absent? No beta-lactamase will be produced. A functional beta-lactamase will be produced. A non-functional beta-lactamase will be produced. Both a functional and a non-functional beta-lactamase will be produced. If the plasmid Lac operon has a mutated promotor that prevents RNA polymerase from binding, which of the following will occur when lactose is present? No beta-lactamase will be produced. A functional beta-lactamase will be produced. A non-functional beta-lactamase will be produced. Both a functional and a non-functional beta-lactamase will be produced.
1. Draw a simplified model of a cis-regulatory element with multiple trans-acting factors up-regulating gene expression. Your simplified model must include: a double-stranded DNA molecule labeled with its appropriate orientation (3' and 5', etc.), an area of DNA highlighting the gene of interest to be transcribed, a highlighted area showing the promoter region, one cis-regulatory element upstream of the promoter with its specific transcription factor • RNApol + a general transcription factor + a TATA binding protein all bound to the appropriate area (think about where these bind before transcription starts), an upstream enhancer with its transcription factor bound to it, • and finally the TSS. All parts must be labeled and in the correct area to earn full credit. **I suggest using different colors for this drawing problem**.
Part A You were sequencing a region of 200 kb in Effrenium voratum (a member of the family Symbiodiniaceae) and you found two genes, an insulator, and two enhancers, as shown below. Gene A Promoter gene A Enhancer I Enhancer I| Insulator Promoter gene b Gene B Q: What would happen if you do some gene editing and you move the insulator between both enhancers? The newly positioned insulator allows enhancer II to activate gene B transcription The newly positioned insulator allows enhancer I to hyperactivate gene B transcription The new positioning of the insulator prevents gene A expression It will be unaltered The newly positioned insulator prevents enhancer II from stimulating the transcription of gene B
Operator | Choose [Choose) A set of genes transcribed together under the regulatory control of a singe promoter A regulatory protein that binds to an region of non-coding DNAgene that suppresses transcriptionby blocking RNA polmer A segment of DNA where the repressor binds to, therety preventing the transcription of certain genes An operon that isalways "off" - turned on by environmental cues an operon that isalways "on"- turned off by environmental cues TChoose Operon Repressor Repressible Operon [ Choose nducible operon [Choose |
amount of protein / amount of mRNA 12 mm / ↑ A B time (minutes) 20 Bacteria were grown in culture. At various time points, substances were added or removed from the culture medium. To assay the effects of the substances on operon gene expression, samples of bacteria were collected at one-minute intervals and levels of specific proteins and mRNAs were determined. If the operon under investigation is a repressible operon, which arrow indicates the addition of a molecule that binds the repressor protein?
Like the lac operon, the hexose operon is controlled by a separate regulatory protein under the control of its own promoter (see the schematic of the operon below). The hexose regulatory protein is sensitive to fatty acyl CoA levels. When all hexose fuel sources are depleted, the bacteria switch to lipid metabolism and fatty acyl CoA levels increase. This turns expression of the hexose operon off. +1 +1 Regulatory Gene P regulator Hexose Operon Genes operon regulator promoter operon promoter e. The regulatory protein that controls expression of the hexose operons is a transcriptional ACTIVTOR or REPRESSOR (circle one).
Match whether the protein binds directly or indirectly (assuming it does) to DNA and whether to the enhancer or promoter. Items may be matched to multiple choices and not all choices will be used. 1. Directly to enhancer Repressors 2. Directly to promoter Activators 3. Indirectly to enhancer TBP associated factors 4. Indirectly to promoter >
MyoD is a transcriptional activator that turns on theexpression of several muscle-specific genes in humancells. The Id gene product inhibits MyoD action.a. One possibility is that the Id protein directly represses the expression of these muscle-specificgenes. Explain how Id would function if it were arepressor.b. Another possibility is that Id inhibits musclespecific gene transcription indirectly, by preventingMyoD function. Explain how Id could function asan indirect repressor.c. Suppose you know the amino acid sequence ofthe Id protein. How might this information supportthe hypothesis in part (a) or in part (b)?
Methylation state of cytosines at the QRT gene promoter is tested in one pair of identical twins, Ana and Becca, at 1 year of age, again when they are 25 years old. At 1 year old, the twins' DNA is the same, and this part of the genome is not methylated. At age 25, Ana's DNA remains unmethylated, but Becca's cells show methylation at this site in skin cells. a. What most likely caused this change in Becca? i. A mutation event ii. An environmental stimulus b. When the twins were 1 year old, QRC mRNA was expressed at similar levels in both Ana and Becca. At age 25, which twin likely expresses more of the QRC mRNA? i. Ana, whose QRC promoter DNA is unmethylated ii. Becca, whose QRC promoter DNA is methylated iii. Both are the same C. At age 25, which twin has more tightly packed chromatin in the region of the QRC gene? Explain briefly. i. Ana, whose QRC promoter DNA is unmethylated ii. Becca, whose QRC promoter DNA is methylated iii. Both are the same d. Some of Becca's skin cells, in…
1. enzymes that catalyze histone acetlation are closesly associated with transription factors, which are proteins that promote transcription. why is this a good biochemical strategy? 2. sp1 is a sqeuence specific human DNA binding protein that binds to a region on the DNA called the GC box, a promoter element with the sequence GGGCGG. Binding of Sp1 to the GC box enhances RNA polymerase 2 activity 50- to a 100 fold. How would you use affinity chromatography to purify SP1?