Concept explainers
a.
To draw:
The recombinant plasmid that indicates the order of the four components and their arrangement in relation to the plasmid vector.
Introduction:
Erythropoietin, which is a human protein stimulates the production of red blood cells. The idea is to make a bacterium that produces this protein, and this recombinant bacterium can be used for the treatment of anemia patients.
b.
To determine:
The element that encodes the ribosome binding site for the mRNA making the fusion protein.
Introduction:
The recombinant plasmid was made to contain coding sequence for human erythropoietin, the regulatory sequence of the lac operon, a sequence encoding E.coli maltose-binding protein, and a sequence encoding a series of five amino acids. These sequences were transformed in the recombinant plasmid to induce the expression of a tagged fusion protein N MBP-DDDDK-erythropoietin C.
c.
To determine:
The elements out of given four that should be placed in the same reading frame with respect to each other.
Introduction:
The four elements present in the recombinant DNA molecule include coding sequence for erythropoietin, a sequence encoding the maltose-binding protein in E.coli and a series of five amino acids, and the regulatory sequence for lac operon.
d.
To determine:
Whether the erythropoietin coding sequences can be obtained from a human genomic DNA or from a human cDNA clone.
Introduction:
The creation of cDNA clones occurs in three major steps known as the synthesis, cloning, and validation. The cDNA clones contain open reading frames and untranslated regions.
e.
To determine:
The compound that could be used for inducing expression of the fusion protein and giving a reason about adding the compound to the medium before seeding with E.coli cells or after reaching the population of cells to high density.
Introduction:
The fusion protein that has to be made in the recombinant E.coli producing erythropoietin protein is N MBP-DDDDK-erythropoietin C.
f.
To determine:
The process by which fusion protein can be purified away from the other E.coli proteins.
Introduction:
The cells that express the fusion protein also contain many other E.coli proteins. It is considered to be important to purify the drugs away from the contaminants. It is known that MBP tightly binds to the sugar maltose and this maltose can be attached to an insoluble resin.
g.
To determine:
The way by which enterokinase can be used to separate the erythropoietin away from the rest of the fusion protein and then perform the purification of the desired pharmaceutical.
Introduction:
It is been studied that the human erythropoietin should not be attached to any other amino acid sequence. So, a protein known as enterokinase is used to cleave the proteins just C-terminal to DDDDK.
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Chapter 16 Solutions
Genetics: From Genes to Genomes
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- Southern blotting is a method used in molecular biology for detection of a specific DNA sequence in DNA samples while northern blotting is used for the detection of RNA in a sample. Write down the similarities and differences between both methods.arrow_forwardCystic fibrosis (CF) is an inherited disorder caused by different types of mutations, many of which prevent ions from moving across cell membranes. Normally there are channel proteins that allow passage of the ions, but in patients with one kind of CF these proteins seem odd. Closer examination shows that these proteins display the correct amino acid sequence. However, they fail to do their job. A) Given that the primary structure of the protein is correct, what can you infer about the DNA sequence for the gene coding this protein on this patient, is there a mutation? Explain. B) Why is the primary structure insufficient to guarantee the proper function of the protein?arrow_forwardListed below are 4 of the 13 genome sites used to create a standard DNA profile. Each site consists of a number of short tandem repeats: sets of 4 nucleotides repeated in a row within the genome. For each site, the number of repeats found at that site for this individual are listed: Imagine you perform a PCR procedure to create a DNA profile for this individual. Which of the following four gels correctly represents the DNA profile of this person?arrow_forward
- In fluorescent Sanger DNA Sequencing, choose one of the following: A):The deoxynucleotides are fluorescently labelled B):Each capillary gel contains a single fluorescent nucleotide C):Chain termination does not occur D):The reaction must be run in the dark E):The incorporated fluorescent nucleotides are detected by a sensor at one end of the capillary gelarrow_forwardA next-generation method of DNA sequencing, 454 sequencing, uses pyrosequencing in which the addition of nucleotides is detected by a:arrow_forwardChoose all of the statements that correctly describe the base pairs drawn below. A C H H-N -H-N N-H- -N B H D موعة Rita N -H---- 2 NHN O- -H-N H -H- N- -H-N The non-Watson-Crick base pair shown in A is much less stable than the base pairs shown in B and C, because the smaller size of the two pyrimidine bases induces a distortion in the structure of the double helix that decreases the stability of the helix when compared to helices with the normal Watson-Crick base pairs. The base pair shown in B is found in BOTH DNA and RNA The base pair shown in C is found ONLY in RNA and NOT DNA The base pair seen in B is more stable than the Watson-Crick base pair shown in C partly because of a larger number of hydrogen bonds and partly because of more favourable pi-stacking interactions with adjacent base pairs.arrow_forward
- Aflatoxin B1 is a highly mutagenic and carcinogenic compound produced by certain fungi that infect crops such as peanuts. Aflatoxin is a large, bulky molecule that chemically bonds to the base guanine (G) to form the aflatoxin-guanine adduct that is pictured below. In the figure below, the aflatoxin is orange, and the guanine base is purple. This adduct distorts the DNA double helix and blocks replication. a. What type(s) of DNA repair system is (are) most likely to be involved in repairing the damage caused by exposure of DNA to aflatoxin B1? b, Recent evidence suggests that the adduct of guanine and aflatoxin B1 can attack the bond that connects it to deoxyribose; this liberates the adducted base, forming an apurinic site. How does this new information change your answer to part (a)?arrow_forwardThe image below shows the base cytosine and a methylated form of cytosine that occurs frequently in the human genome. Use your knowledge of DNA structure to answer the following questions: a) Does methylation of cytosine affect its ability to base-pair with guanine? Explain your answer. b) Would methylation of cytosine affect the binding of a protein that interacts with a C-G base-pair in the major groove?arrow_forwarda) Explain the effect of the guanine:cytosine ratio on melting temperature of DNA. b) The Hershey-Chase experiment proved that DNA is the genetic material and not protein. Explain in detatil how this experiment was conducted.arrow_forward
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