iii,iv ,v Clary Fray used the pET vector system to express her prokaryotic amylase enzyme. She addedpeptone into her culture broth of BL21(DE3) Escherichia coli strain to induce proteinexpression. At the end of the experiment, she discovered that her protein was not expressed.She repeated three more times but her protein of interest was still not produced.(i)(ii)(iii)(iv)(v)Explain the reason why Clary failed to obtain her protein of interest and suggest asolution to troubleshoot this problem.Clary plans to express her protein along with a polyhistidine-tag. Explain theimportance of His-tag in protein work.Is DH5a Escherichia coli suitable to propagate the plasmid before protein expression?Besides heat shock method, elaborate another procedure Clary could utilize totransform the recombinant pET vector into the host cell.If her supervisor instructs her to express a gene from gold fish (Carassius auratus), isthe expression system she is using now suitable for this experiment? Explain.

Name: iii,iv ,v Clary Fray used the pET vector system to express her prokary
Uploaded: 2024-03-20T06:42:55.000Z
Channel: Bartleby
Description: iii,iv ,v Clary Fray used the pET vector system to express her prokaryotic amylase enzyme. She addedpeptone into her culture broth of BL21(DE3) Escherichia coli strain to induce proteinexpression. At the end of the experiment, she discovered that her

i). The protein expression in the pET vector cultured in BL21(DE3). The IPTG induction at specific…

Answered: iii,iv ,v

Science

Biology

iii,iv ,v

Human Heredity: Principles and Issues (MindTap Course List)

11th Edition

ISBN:9781305251052

Author:Michael Cummings

Publisher:Michael Cummings

Chapter10: From Proteins To Phenotypes

Section: Chapter Questions

Problem 20QP: If an extra nucleotide is inserted in the first exon of the beta globin gene, what effect will it...

See similar textbooks

Similar questions

In studies of the amino acid sequence of wild-type and mutant forms of tryptophan synthetase in E. coli, the following changes have been observed: Determine a set of triplet codes in which only a single-nucleotide change produces each amino acid change.
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?
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?
If an extra nucleotide is inserted in the first exon of the beta globin gene, what effect will it have on the amino acid sequence of the globin polypeptides? Will the globin most likely be fully functional, partly functional, or nonfunctional? Why?
The protein encoded by the cystic fibrosis gene is 1480amino acids long, yet the gene spans 250 kb. How is thisdifference possible?
Most of the mutations that Yanofsky recovered were missense mutations. However, Yanofsky also recovered a nonsense mutation that changed amino acid number 15 into a stop codon. This codon normally encodes Lysine. Does the recovery of this mutation support the hypothesis that this Lysine residue is critical in the function of the tryptophan synthetase protein? Why or why not?
In a line of human cells grown in culture, a geneticist isolates a temperature-sensitive mutation at a locus that encodes an acetyltransferase enzyme; at temperatures above 38°C, the mutant cells produce a nonfunctional form of the enzyme. What would be the most likely effect of this mutation if the cells were grown at 40°C?
Based on the following wild type sequence, indicate if each of the mutations should be classified as : insertion, deletion, missense, nonsense, silent (Use the provided Genetic Code table). Wild Type: GUC GCC GAC GAG AGG Mutant 1: GUC GCC AGA CGA GAG G Mutant 2: GUC GCC ACG AGA GG Mutant 3: GUC GCC AAC GAG AGG Mutant 4: GUA GCC GAC GAG AGG Mutant 5: GUC GCC GAC UAG AGG
What is known as catabolite repression ?
As part of a project investigating potential new drug targets in the fight against malaria, you are seeking to clone the gene for a protein from the malaria parasite Plasmodium falciparum. You wish to express this protein in BL21 (DE3) cells, a standard laboratory strain of Escherichia coli. After purification of your protein, you run an SDS-PAGE gel and notice that the major band has lower molecular weight than expected, so you fear you are getting a truncated version. 1. What technique could you use to confirm that you are obtaining a shortened version of your intended protein? explain
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 +
You are interested in studying resistance to heavy metals and have selected the yeast Saccharomyces cerevisea to conduct your studies. You have recovered a deletion mutant that does not tolerate high concentrations of zinc (grows poorly in zinc containing media ) and have designated the mutant pgz-1 (for poor growth in zinc ). (a) What is the advantage to the type of mutant used in this work? What class of mutagen was likely use to generate pgz-1? ( b) Do you expect the PGZ gene to be expressed in your mutant? Explain.