Zoey Wong is a research officer at the Department of Biosciences of Tunku Abdul RahmanUniversity College. Her supervisor instructs her to prepare chemically competent cells forheat shock transformation from old batches of competent cells that are already available in thefreezers. The competent cells will eventually be used for the expression of a prokaryoticenzyme using the pET vector system.(i)(ii)You found several different strains of Escherichia coli in the -80 C freezer. Statethree (3) E. coli strains you would use for your project which involves the cloningand expression of the protein of interest.In order for cloning and expression to happen optimally, suggest two (2) types of pETvectors suitable for your project.

The basic principles of the central dogma of molecular biology is similar in both prokaryotic and…

Answered: Zoey Wong is a research officer at the…

Biochemistry

9th Edition

ISBN:9781319114671

Author:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.

Publisher:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.

Chapter1: Biochemistry: An Evolving Science

Section: Chapter Questions

Problem 1P

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The following DNA sequence is from a bacteriophage that infects a pathogenic bacterium and scientists want to know if this bacteriophage could prove to be a potential treatment against it. But first scientists need to discover if different strains of this pathogen have restriction endonucleases that it may use for its own protection. They try 3 different RE’s:a) EcoR1 b) HaeIII c) BamH1 Look up the recognition sequences for the 3 Res. Enzymes above and check whether the phage genome (a snippet of which is shown below) will or will not be ‘cut’. Tell me how their experiment worked out and what their conclusion was.G A A A A G G C C A C A A G G C C G T C G A C T T T T A A A A G G C C A C A T G C G G C T T T T C C G G T G T T C C G G C AG C T GA A A AT T T T C C G G T G T A C G CC
We have two specific strains of E. coli that have shown horizontal gene transfer (HGT) when mixed. To experimentally determine the method of HGT that is happening, the following conditions are set up in different tubes of culture media: A) Donor and recipient strain mixed together (control - no treatment). B) Donor and recipient strains mixed together, DNase added (can digest DNA in solution, not within cells).C) Special tube containing a membrane filter (with pores that allow DNA and viruses to pass through, but not bacterial cells) that separates two compartments. Donor strain is added on one side, the recipient strain on the other (they are separated by the filter).D) Donor and recipient strains mixed together, with chemical that inactivates viruses (chemical affects bacteriophages in solution so they are unable to attach to cells). The results: Tubes A, B, and D: HGT was observed. Tube C: HGT was NOT observed. Based on this, which type of HGT was occurring? Conjugation,…
A research group is studying a bacterium X that binds to mucosal cells in the lung and invades. Wildtype X has an LD50 value of 10 bacteria when administered to mice by inhalation. Using transposon mutagenesis, the researchers have isolated two mutants of X that they call Xmut1 and Xmut2, both of which have LD50 values of 105 when inhaled by mice. However, in tissue culture cells, Xmut1 can invade the cells just as well as wild-type X, while Xmut2 cannot. Provide a possible explanation for these results.
In 1944, Avery, Macleod, and McCarty provided strong evidence that DNA is the hereditary material in Streptococcus pneumoniae by Group of answer choices showing that avirulent cells could become virulent by the process of transduction none of these is true. showing that virulent cells could become avirulent if the DNA was destroyed after transformation showing that avirulent cells could not gain the ability to become virulent cells if conjugation was interrupted. showing that avirulent cells could not gain the ability to become virulent if DNA was destroyed after transformation.
A pure culture of an unknown bacterium was streaked onto plates of a variety of media. You notice that the colony morphologyis strikingly different on plates of minimal media with glucose compared to that seen on trypticase soy agar plates. How can you explain these differences in colony morphology? Also, describe what happens when a nonsense mutation is introduced into the gene encoding transposase within a transposon and why is it more likely that insertions or deletions will be more detrimental to a cell than point mutations?
Identify the most mistaken (wrong) choice: a) Transcription machinery and an enhancer can bind to the chromosome at the same time. b) Organic matters may interfere with heat treatment of bacterial growth control. ( c) Nitrocellulose can be used to filter out microorganisms from a liquid solution. d) Time to kill a bacterial culture is not proportional to the number of microbes in the culture
Regarding the cultivation of animal cells in vitro, it is correct to state that: *(Only one statement is correct) a) Transformed cells differ from normal and established cells in that they have the ability to proliferate indefinitely in culture b) Transformed cells derive from isolated and dissected tumors, constituting cell lines that have limited cultivation time c) The genetic instability presented in normal cells makes them preferential models for the production of recombinant proteins d) Cells cultured in adhesion must be periodically expanded, through the use of enzymes such as trypsin e) The metabolism of carbon sources in the culture medium by animal cells generates products such as lactate, which positively influence their productivity
Ahmed had isolated bacteria from the sputum of a patient that seemed somewhat resistant to their antibiotic treatment, since multiple weeks of treatment had yielded no satisfactory improvement of their active disease. The researcher had then resuspended the patient's sputum in saline and used a mouse model to measure the efficiency of a phage selected for its capacity to target M. mycobacterium, as a single therapy, and as a combination therapy with antibiotics. Here is a schematic representation of the experiment: Group 1 Control (saline) Inoculate mice with patient sputum Group 2 wait for active disease to show Treated with phage Sputum samples Collect lung sample every 2 weeks Tuberculosis patient Group 3 Treated with Abx Split mice showing active disease in 4 Group 4 Treated with Abx and Count bacteria in sample groups phage Figure 4. Sputum from patients were isolated and resuspended in saline. Mice were given equal amounts of resuspended sputum intra-nasally and were left to…
Ahmed had isolated bacteria from the sputum of a patient that seemed somewhat resistant to their antibiotic treatment, since multiple weeks of treatment had yielded no satisfactory improvement of their active disease. The researcher had then resuspended the patient's sputum in saline and used a mouse model to measure the efficiency of a phage selected for its capacity to target M. mycobacterium, as a single therapy, and as a combination therapy with antibiotics. Here is a schematic representation of the experiment: Group 1 Control (saline) Inoculate mice with patient sputum Group 2 wait for active disease to show Treated with phage Sputum samples Collect lung sample every 2 weeks Tuberculosis patient Group 3 Treated with Abx Split mice showing active disease in 4 Group 4 Treated with Abx and Count bacteria in sample groups phage Figure 4. Sputum from patients were isolated and resuspended in saline. Mice were given equal amounts of resuspended sputum intra-nasally and were left to…
Ahmed had isolated bacteria from the sputum of a patient that seemed somewhat resistant to their antibiotic treatment, since multiple weeks of treatment had yielded no satisfactory improvement of their active disease. The researcher had then resuspended the patient's sputum in saline and used a mouse model to measure the efficiency of a phage selected for its capacity to target M. mycobacterium, as a single therapy, and as a combination therapy with antibiotics. Here is a schematic representation of the experiment: Group 1 Control (saline) Inoculate mice with patient sputum Group 2 wait for active Treated with phage disease to show Collect lung sample every 2 weeks Tuberculosis Sputum samples patient Group 3 Treated with Abx Group 4 Split mice showing active disease in 4 Treated with Abx and Count bacteria in sample groups phage Figure 4. Sputum from patients were isolated and resuspended in saline. Mice were given equal amounts of resuspended sputum intra-nasally and were left to…
The Pfizer Covid-19 vaccine is one of the vaccines currently being rolled out for mass vaccination in South Africa to protect the population against SARS CoV-2, and the active compound in this vaccine is an mRNA molecule. a) The starting material for productio of this vaccine is a copy of the coronavirus spike protein gene cloned into a ppasmid. List the main experimental steps in the vaccine production process, to go from a plamid to an mRNA molecule. b) Which molecular processes in the cell will allow an injected strand of mRNA to produce anti-Covid immunity in the vaccine recipient?
Bacteriophage P22 was used in generalized transduction experiments to infect the Salmonella typhimurium donor strains described in the table below. The resulting phage lysates were then used to infect the S. typhimurium recipient strains listed in the table. In each cross, a phenotype was selected for one of the three genetic markers studied (str, aceA, thrA), and then replicates were performed to select the corresponding recombinants for the other two markers. The results are given in the following table: Recipient strain Selected phenotype Selected recombinants Donor strain str thrA aceA+ thrA str aceA+ strs thrA+ aceA thrA+ str aceA Str Ace+ Str ThrA ThrA+ ThrA ThrA+ Ace Ace str: gene involved in streptomycin resistance, aceA gene involved in the use of acetate as a carbon source, thrA: gene involved in the biosynthesis of threonine. Number 60 40 95 5 10 90 Determine the order of the genes and draw a genetic map showing this order

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