DNA can be cut at specific sequence motifs using enzymes called restriction endonucleases. You obtain a single PCR product from R. sphaeroides DNA that can be cut in the places shown below by solid arrows. These are the Clal specific restriction motifs. Positions of these restriction sites are indicated by nucleotide numbers in a 1023 nucleotide DNA fragment. Draw the expected band(s) for each of the bacterial species on the gel picture below if treated with the restriction enzymes as illustrated. Rhodobacter sphaeroides 2. 4. 1 0. 1023 150 627 Rhodopseudomonas palustris BisB5 0. 1023 627 Dinoroseobacter shibae DFL 12 1023 150 Silicibacter sp. TM 1040 1023 DNA ladder R sphaeroides 2. 4. 1 R palustris BisB5 D. shibae DFL12 Silicibacter sp. TM1040 1100 1000 900 800 700 600 500 400 300 200 100

DNA can be cut at specific sequence motifs using enzymes called restriction endonucleases. You obtain a single PCR product from R. sphaeroides DNA that can be cut in the places shown below by solid arrows. These are the Clal specific restriction motifs. Positions of these restriction sites are indicated by nucleotide numbers in a 1023 nucleotide DNA fragment. Draw the expected band(s) for each of the bacterial species on the gel picture below if treated with the restriction enzymes as illustrated. Rhodobacter sphaeroides 2. 4. 1 0. 1023 150 627 Rhodopseudomonas palustris BisB5 0. 1023 627 Dinoroseobacter shibae DFL 12 1023 150 Silicibacter sp. TM 1040 1023 DNA ladder R sphaeroides 2. 4. 1 R palustris BisB5 D. shibae DFL12 Silicibacter sp. TM1040 1100 1000 900 800 700 600 500 400 300 200 100

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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b. After molecular cloning and confirming you properly cloned stripes into the vector shown, you transform bacteria with the plasmid and select out bacteria that contain the plasmid. Now, you want to these transformed bacteria to express the STRIPES. Under what conditions would we need to grow bacteria transformed with this plasmid so that they express STRIPES? Explain your answer, and be sure to describe the important regulatory regions on the plasmid above and what interacts with these regulatory regions in the conditions you have indicated.
A 2.0kb bacterial plasmid ‘BS1030’ is digested with the restriction endonuclease Sau3A; the plasmid map is depicted in the diagram below and the Sau3A (S) restriction sites are indicated. Which of the following DNA fragments do you expect to see on an agarose gel when you run Sau3A-digested plasmid ‘BS1030’ DNA? a. 250 bp, 450 bp, 550 bp, 1.1 kb, 1.5 kb and 2.0 kb b. 2.0kb c. 250 bp, 400 bp, 450 bp, 500 bp and 550 bp d. 100 bp, 200 bp, 250 bp, 400 bp, 500 bp and 550 bp
Draw a diagram showing what pGEM will look like after it has been digested with BamHI. Be sure to show both strands (with enzyme sequence) and include the origin of replication, the ampicillin resistance gene, 3’ and 5’, and the DNA sequence at the restriction site. It will probably be easiest to simply draw it as an open circle, not a line. Now show what pGEM will look like after it has been digested with EcoRI instead of BamHI. Again, be sure to show both strands and include the origin of replication, the ampicillin resistance gene, 3’ and 5’, and the DNA sequence at the restriction site.
a. How many restriction fragments did the digest produce in Lane 3? What does that mean about the number of restriction sites that species had in its DNA? b. Provide your estimates for the lengths of the restriction bands in Lane 3.
Which of the following sequences, in the double-stranded form, can probably be recognized by a restriction enzyme? Choose an answer from below? TTCCTT CGACGA GTCGAC GGTTGG GAGGAC
Calculate the volume (in µL) of DNA template required to add 60.0 ng to a 40.0 µL PCR if the DNA template is at a concentration of 28.0 ng µL-1.
Restriction endonucleases are bacterial enzymes that cleave duplex (double-stranded) DNA at specific nucleotide sequences. The mode of replication of the animal virus SV40 has been investigated by using restriction endonucleases that cleave SV40 DNA into a number of unique segments. Like most viruses, SV40 DNA is circular. The map positions of the 11 fragments produced by a pair of restriction endonucleases are shown on the next page. Immediately following a 5 or 10 minute pulse of radioactively labeled thymidine, labeled SV40 molecules that have completed replication during the pulse are isolated. These newly replicated DNA molecules are digested by the restriction endonucleases and the resulting fragments are analyzed for the relative amounts of pulse label they contain. The results are in the table below. Assume that at the time the label was added there was a random population of replicating SV40 DNA molecules in all possible stages of synthesis. From the information given below,…
A linear DNA fragment was produced by digestion with the restriction enzyme, Xba1. This fragment with XbaI(X) sites on both ends was then further digested with HindIII (H) and EcoRI (E). Draw a restriction map of the linear fragment based on the gel electrophoresis results shown below. X H Marker E H/E __2000bp __ __1500b __1300bp __ __ __1000bp __ __700bp __ __500bp __400bp __300bp __ __200bp __ __ __100bp
Consider the ends of the DNA fragments shown below. They have been produced by digestion of a single sequence of DNA using a number of restriction endonucleases. 1. 5'A 3' 3'TTCGA5' 2. 5'G 3' 3'CAGCT5' 3. 5'AATTC3' 3' G5 4. 5'TCGAC3' 3' G5' 5. 5'GGG 3' 3'CCC 5' Which of these ends are capable of annealing and being joined by DNA ligase?
Use the plasmid map for pBashi below to answer the following questions Xhol Hind!!! 15 kb 11 kb 10 kb 8 kb 7 kb 5 kb a. How large is pBashi? b. On the gel below, draw the bands corresponding to the expected pBashi cleavage products after restriction digest with the indicated restriction enzymes (Lane 2 - Xhol only, Lane 3 - HindIII only, Lane 4 - Xhol+ HindIII) 4 kb 3 kb 3 kb 2 kb 1 kb 4 kb Hindill 1 kb 7 kb Xhol Xhol Xhol Xhol Hind!!! Ladder Xhol Hindill Hind!!! Pstl Pstl c. Lane 5 in the gel above displays the pBashi cleavage products after restriction digest with the restriction enzymes Xhol+ Pstl. Lane 6 in the gel above displays the pBashi cleavage products after restriction digest with the restriction enzymes Xhol+ HindIII + Pstl. Indicate your final answers to the questions below clearly on the pBashi plasmid above; you will NOT receive any credit if you only list sizes of fragments below. If you need to, draft your answer somewhere else before writing your final answer above.…