Concept explainers
A
a. What length of DNA is bound by the transcriptional proteins? Explain how the gel results support this interpretation.
b. Draw a diagram of this DNA fragment bound by the transcriptional proteins, showing the approximate position of proteins along the fragment. Use the illustration style seen in Research Technique
c. Explain the role of DNase I.
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GENETIC ANALYSIS: INTEGRATED - ACCESS
- Clone number in this case is number 196 which is shown in the images. State whether a BamHI site has been re-created at the forward- and the reverse-end junctions of the human DNA with the plasmid vector band sizes are shown in one of the images. (0.5, 1, 2, 3, 4, 5, 6, 8, 10kb)arrow_forwardMass spectrometry is a powerful tool in proteomics. What are the four key features of a mass spectrometer? Describe briefly how MALDI and two-dimensional polyacrylamide gel electrophoresis could be used to identify a protein expressed in cancer cells but not in normal healthy cells.arrow_forwardWhat are DNA probes Describe three (3) methods that can be used to label DNA probes Describe 2 methods for detection of the hybridized probe List the advantages of using non-radioactively labeled probes vs radioactively labeled probes.arrow_forward
- Expressing genes encoding a particular protein’s subunits from a plasmid in E. coli cells (volume of 1 cell = 0.5 um3) results in 1 gram of this protein when the E. coli cells reach a density of 2 x 109 cells/mL in a culture volume of 1.0L. Is this possible? Why or why not?.arrow_forwardThe cellulase gene of Bacillus licheniformis was successfully cloned into the pET21a vector and expressed in Escherichia coli BL21. The pET21a vector consists of ampicillin resistant gene. To screen for the successful transformants, E. coli BL21 was cultivated on LB agar containing ampicillin (100 pg/mL) and 0.5% (w/v) carboxymethylcellulose, and incubated at 37 0C for 6 hours. After that the agar plate was stained with Congo red solution for 15 minutes and washed twice with sodium chloride solution and the observation is as shown in Figure 3. Answer the following: (i.) Briefly explain why ampicillin was added to the LB agar. (ii) Indicate the function of carboxymethylcellulose in the LB agar. (iii) Conclude how the researchers were able to identify the E. coli BL21 that carried the cellulase gene.arrow_forwardYou observe the following: Your Your friend's Gel Gel 120 kDa 100 kDa 120 kDa 100 kDa -80 kDa 80 kDa -40 kDa 40 kDa a. You and your friend analyzed the same protein sample in your gels. Why are your results different? Explain briefly which gel is the correct representation of the SDS-PAGE & Western blot, and why? b. Explain briefly the size of protein observed in the chromatography column and the Western blot. Are they different or the same, why? (S c. If the primary antibody to Protein A was made in a mouse, describe the details of the secondary antibody used in the Western blot analysis?arrow_forward
- These are the results after running Agarose Gel Electrophoresis of the cut/uncut pUC19 plasmid and isolated Genomic DNA at 120 V for 30 minutes. Can you explain what this gel means?arrow_forwardIn Northern blot analysis, mRNA samples from tissues are bound to a labeled DNA probe that is complementary to the mRNA, and run on a gel to be visualized. The protein tropomyosin is known to be present in both brain and liver. When brain and liver tissue were assayed for the presence of tropomyosin mature mRNA, bands of two different sizes were seen. Tropomyosin gene diagram (3000 bp total): Shown in attatched image If the band on the Northern blot for mRNA isolated from liver tissue is 2580 bp, whereas from brain tissue the band is 2250 bp, what is most likely? a)The two mRNAs are made from different tropomyosin DNA sequences. b)Exon 2 is alternatively spliced out of the brain mRNA. c)Introns 1 and 2 are spliced out of the brain transcript but not the liver transcript. d)Exons 1 and 3 are spliced out of the brain transcript but not the liver transcript. e)Exon 2 is alternatively spliced out of the liver mRNA.arrow_forwardWhat volume of 4X loading buffer must be added to 21 micro L of DNA in a technique for DNA sample preparation for agarose gel electrophoresis to generate a 1X buffer solution?arrow_forward
- Analysis of a 49kDA protein is aimed with a western blot technique. For this purpose, "whole cell extract" was obtained from biopsy samples taken from three different individuals, and when their spectrophotometric measurements were made, their concentrations were determined to be 40 µg/µL, 50 µg/µL, and 100 µg/µL for three samples, respectively. At the beginning of the experiment, 200 µg of protein is loaded into each gel well by using a ready 5x loading solution containing SDS and β-mercapta-ethanol together with glycerol and methylene blue dye. a) If loading will be done in a volume of 20µL, then how is the preparation of the samples before loading?b) Considering the loading content given above, what can be said about whether this PAGE is "non-denaturing" or "denaturing". Justify taking into account the loading solution content.arrow_forwardHow much 340bp insert DNA should be added to a ligation in which 50ng of 4.3kb vector will be used? The desired vector: insert molar ratio needs to be 1:3.arrow_forwardA cloned Bam HI DNA fragment, 3.0 kb long, was isolated in preparation for sequence determination. First, a map of restriction endonuclease cleavage sites had to be prepared. Cleavage by each of the indicated enzymes generated fragments of the sizes shown on the three agarose electrophoresis gel figures below; the gels are running from top to bottom. On the diagram below those, show the relative positions of EcoR I and Hpa II cleavage sites and the distances (in kb) between them. Explain your reasoning – is your placement of the cleavage sites the only one that will work?arrow_forward
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