Mabelle used the pET vector system to express her prokaryotic amylase enzyme. She addedIPTG into her culture broth of DH5a Escherichia coli strain. At the end of the experiment,she discovered that her protein was not expressed. She repeated three more times but herprotein of interest was still not produced.(i)(ii)Explain the reason why Mabelle failed to obtain her protein of interest and suggest asolution to troubleshoot this problem.Mabelle plans to express her protein fused to a polyhistidine-tag (His-tag). Explainthe importance of His-tag in protein work.

In expression and purification of protein from bacterial system, the following steps are performed:…

Answered: Mabelle used the pET vector system to…

Biology: The Dynamic Science (MindTap Course List)

4th Edition

ISBN:9781305389892

Author:Peter J. Russell, Paul E. Hertz, Beverly McMillan

Publisher:Peter J. Russell, Paul E. Hertz, Beverly McMillan

Chapter15: From Dna To Protein

Section15.1: The Connection Between Dna, Rna, And Protein

Problem 1SB

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Given the following phenotypes, explain how the mutation (identified by a (-) superscript) will affect the E. coli grown in lactose medium. a. l+p+o+z-y+ b. i-p+o+z+y+ c. l+p+o-z+y+ d. l+p-o+z+y+
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?
Given the following phenotypes, explain how the mutation (identified by a (-) superscript) will affect the E. coli grown in lactose medium. l+p+o+z-y+ i-p+o+z+y+ l+p+o-z+y+ l+p-o+z+y+
Given the following genotypes, explain, by answering the questions in each number, how the mutation (identified by a (-) superscript) will affect E. coli grown in lactose medium. Will there be a complete set ofgene products? (Yes/No) Will the lac operon be turnedon/off? Will the cell survive? (Yes/No) a. i + p + o + z - y + b. i + p - o + z + y + c. i + p + o - z + y +
For each of the E. coli strains containing the lacoperon alleles listed, indicate whether the strain isinducible, constitutive, or unable to expressβ-galactosidase and permease.a. I+ o+ Z− Y+/ I+ ocZ+ Y+b. I+ o+ Z+ Y+/ I− ocZ+ Y−c. I+ o+ Z− Y+/ I− ocZ+ Y−d. I−P− o+ Z+ Y−/ I+ P+ ocZ− Y+e. Iso+ Z+ Y+/ I− o+ Z+ Y−
In E. coli, the gene bioD+ encodes an enzyme involved in biotin synthesis, and galK+ encodes an enzyme involved in galactose utilization. An E. coli strain that contained wild-type versions of both genes was infected with P1 phage, and then a P1 lysate was obtained. This lysate was used totransduce (infect) a strain that was bioD− and galK−. The cellswere plated on a medium containing galactose as the sole carbonsource for growth to select for transduction of the galK+ gene.This medium also was supplemented with biotin. The resultingcolonies were then restreaked on a medium that lacked biotin tosee if the bioD+ gene had been cotransduced. The following resultswere obtained:What information do you know based onthe question and your understanding of the topic?
In E. coli, the gene bioD+ encodes an enzyme involved in biotin synthesis, and galK+ encodes an enzyme involved in galactose utilization. An E. coli strain that contained wild-type versions of both genes was infected with P1 phage, and then a P1 lysate was obtained. This lysate was used totransduce (infect) a strain that was bioD− and galK−. The cellswere plated on a medium containing galactose as the sole carbonsource for growth to select for transduction of the galK+ gene.This medium also was supplemented with biotin. The resultingcolonies were then restreaked on a medium that lacked biotin tosee if the bioD+ gene had been cotransduced. The following resultswere obtained:What topic in genetics does this question address?
In Hershey-Chase experiment, bacteriophages protein coats were tagged with radioactive isotope S-32. These phages were used to infect E. coli cells and the cells were further centrifuged to form pellets. Why was the radioactivity level of S-32 found greater outside the cells compared to the E. coli cell pellets? Explain briefly. If the experiment is repeated in the same manner but this time the phage protein coats are labelled with isotope X and the phage DNA with isotope Y, which isotope’s radioactivity will be found in greater amounts in the E. coli cell pellets after centrifugation? Explain briefly.
In the procedure shown, why was it necessary to link thecoding sequence for the A or B chains to the sequence forβ-galactosidase? How were the A or B chains separated fromβ-galactosidase after the fusion protein was synthesized in E. coli?
Refer to the diagram of pUC18 (Fig.) to determine which restrictionenzymes you could use to insert a gene that would not interfere with ampicillin resistance or production of β-galactosidase by the host cell.
What are the reagents and materials used/needed in the extraction of invertase from yeast? for a lab experiment regarding enzymes.
When Avery and his colleagues had obtained what was concluded to be the transforming factor from the IIIS virulent cells, they treated the fraction with proteases, RNase, and DNase, followed in each case by the assay for retention or loss of transforming ability. What were the purpose and results of these experiments? What conclusions were drawn?

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