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.
Trending nowThis is a popular solution!
Chapter 16 Solutions
Genetics: From Genes to Genomes
- Ethanol (CH3-CH2-OH) is miscible in water because it is able to form hydrogen bonds with itself and other molecules. However, its structure only allows it to form 1-2 hydrogen bonds. This is one reason why even low concentrations of ethanol in solution are lethal for cells. Based on this information, explain why we can use high concentrations of ethanol to precipitate DNA out of solution. Also, describe/predict the effects of increasing concentrations of ethanol in (and around) a cell on macro-molecular interactions (i.e. on weak bonds). Finally, it is possible to select for yeast that are tolerant to increased concentrations of ethanol. Give an example of a physiological change in yeast cells that might make them resistant to ethanol.arrow_forwardA compact disc (CD) stores about 4.8 × 109 bits of information in a 96 cm2 area. This information is stored as a binary code—that is, every bit is either a 0 or a 1. how many bits would it take to specify each nucleotide pair in a DNA sequence? how many CDs would it take to store the information contained in the human genome?arrow_forwardAilee is interested to determine the nucleotide sequence of her bacterial heat shock gene. Hence, DNA sequencing needs to be performed for this analysis. One of the earliest methods invented is known as Sanger sequencing. Explain in detail the mechanism of this sequencing technique with the aid of a simple diagram.arrow_forward
- 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_forwardAn example sequence corresponds to human sickle cell beta-globin mRNA and that this disease results from a point mutation in the β globin gene. In the following section, you will compare sickle cell and normal β globin sequences to reveal the nature of the sickle cell mutation at the protein level. To do this you need to find at least one sequence representing the normal beta globin gene. Open a new window and visit the NCBI home page(http://www.ncbi.nlm.nih.gov) and select “Nucleotide” from the drop menu associated with the top search box. Then enter the search term: HBB . Note that lots of irrelevant results are returned so let's apply some “Filters” (available by clicking in the left-hand sidebar) to focus on RefSeq entries for Homo sapiens. Remember that we are after mRNA so we can compare to the mRNA sequence from part 1 above. QUESTION #1: What is the ACCESSION number of the “Homo sapiens hemoglobin, beta (HBB), mRNA” entry? NOTE: Boolean operators (NOT, AND, OR) as…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_forward
- Listed 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_forwardIn 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_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_forwardYou have sequenced the genome of the bacterium Salmonella typhimurium, and you are using BLAST analysis to identify similarities within the S. typhimurium genome to known proteins. You find a protein that is 100 percent identical in the bacterium Escherichia coli. When you compare nucleotide sequences of the S. typhimurium and E. coli genes, you find that their nucleotide sequences are only 87 percent identical.a. Explain this observation.b. What do these observations tell you about the merits of nucleotide- versus protein-similarity searches in identifying related genes?arrow_forward
- Human Anatomy & Physiology (11th Edition)BiologyISBN:9780134580999Author:Elaine N. Marieb, Katja N. HoehnPublisher:PEARSONBiology 2eBiologyISBN:9781947172517Author:Matthew Douglas, Jung Choi, Mary Ann ClarkPublisher:OpenStaxAnatomy & PhysiologyBiologyISBN:9781259398629Author:McKinley, Michael P., O'loughlin, Valerie Dean, Bidle, Theresa StouterPublisher:Mcgraw Hill Education,
- Molecular Biology of the Cell (Sixth Edition)BiologyISBN:9780815344322Author:Bruce Alberts, Alexander D. Johnson, Julian Lewis, David Morgan, Martin Raff, Keith Roberts, Peter WalterPublisher:W. W. Norton & CompanyLaboratory Manual For Human Anatomy & PhysiologyBiologyISBN:9781260159363Author:Martin, Terry R., Prentice-craver, CynthiaPublisher:McGraw-Hill Publishing Co.Inquiry Into Life (16th Edition)BiologyISBN:9781260231700Author:Sylvia S. Mader, Michael WindelspechtPublisher:McGraw Hill Education