In this western blot, the levels of actin increase with increasing amounts/lexpression of the viral protein (VP). Figure description; Increasing amounts of a plasmid expressing the viral protein (0.5, 1, or 2ug) were cotransfected with TBK1 expression plasmid. Cells were harvested 24 h post-transfection and analyzed for phosphorylated TBK1 (anti-PTBK1 Ser172), total TBK1 (anti-TBK1), B-actin (anti-B-actin), and viral protein (anti-VP) expression by Western blot analysis. VP PTBK1 (S172) TBK1 actin Virus proteins O True O False
Q: Imagine that you completed your ligation of the gRNA insert (annealed oligos) into cut pML104 vector…
A: Nuclear localization signals (NLS) is importnat because it helps to transport Cas9 into the nucleus…
Q: You observe the following: Your Your friend's Gel Gel 120 kDa 100 kDa 120 kDa -100 kDa -80 kDa 80…
A: Blotting, as we all know, is a process that uses electrophoresis to separate DNA, RNA, and protein…
Q: T. Miyake and M. Demerec examined proline-requiring mutations in the bacterium Salmonella…
A: In transduction process, the closer the genes are present on a DNA, the more possibility they have…
Q: Which of the following set(s) of primers a-d could you use to amplify the following target DNA…
A: DNA is the genetic material in most living organisms. It is the information hub of the cell that…
Q: Below is a diagram of the vector you are planning to use. You identify four restriction enzyme…
A: Restriction enzymes are the cleaving enzymes and thus are also known as molecular scissors. These…
Q: You are cloning the genome of a new DNA virus into pUC18. You plate out your transformants on…
A: Molecular cloning is a technique by which a gene of interest is inserted into a plasmid vector,…
Q: In the Western blot shown here, proteins were isolated from redblood cells and muscle cells from two…
A: Thalassemia is a blood disorder passed down through families (inherited) in which the body makes an…
Q: A researcher has a plasmid vector that has been cut with a restriction enzyme that cuts after the…
A: Restriction enzymes are also called "restriction endonucleases". They are a group of enzymes that…
Q: he bands indicated by arrow "A" (to the left of the western blot) most likely represent: A.…
A: The bands indicated by arrow "A" (to the left of the western blot) most likely represent:-
Q: A plasmid that is both ampicillin and tetracyclineresistant is cleaved with PstI, which cleaves…
A: A plasmid is an extrachromosomal small DNA molecule present in a cell and is also physically…
Q: In this western blot, the levels of TBK remain constant with increasing amounts/expression of the…
A: TBK-1 gene encodes an enzyme known as TBK-1 (TANK binding kinase-1). This is a…
Q: In this western blot, the levels of phosphorylated TBK (PTBK) decrease with increasing…
A: Western blot analysis is the assay done to check expression of protein. In this method, proteins are…
Q: Can you answer the last three q
A: Note: As per Bartleby Guidelines For Remaining Answers Please Repost The Question. Introduction:…
Q: When cloning a foreign DNA fragment into a plasmid, it is often useful to insert the fragment at a…
A: Recombinant DNA technology is the modern technology that is used to engineer DNA molecules that have…
Q: E. coli strains diploid for the lac region were constructed by introducing a plasmid carrying the…
A: ANSWER;- I- mutation- repressor is unable to bind to the operator region. If there is no other…
Q: In E. coli, the gene bioD+ encodes an enzyme involved in biotin synthesis, and galK+ encodes an…
A: Transduction is a process of gene transfer in which DNA (deoxyribonucleic acid) from a source is…
Q: How does the insertional inactivation ofb-galactosidase allow the presence of foreign DNAin a…
A: Insertional Inactivation: it is a method of recombinant DNA technology , which is employed to pick…
Q: To estimate the number of cleavage sites in a particular piece of DNA with a known size, you can…
A: DNA= deoxyribonucleic acid and this is found in the mitochondria of cells and is encoded in the…
Q: A Western blot is seen in the picture above. Sodium dodecyl sulfate gel polyacrylamide gel…
A: The western blot, also known as western blotting, is a molecular biology and immunogenetics…
Q: The genotypes of five merozygote bacteria are shown below. In each set, the enotype of the lac…
A: operon Operon can be defined as a multiple gene that have only single promoter. All these genes are…
Q: The diagram below shows that of a plasmid containing genes that confers erythromycin (Erm®) and…
A: Plasmids are small circular double-stranded DNA molecules that store additional genetic information…
Q: Where is the F plasmid located in an Hfr cell? Where is the F plasmid located in an F+ cell?
A: Prokaryotic cells are unicellular organisms that differ from multicellular organisms in composition…
Q: For each genotype in the table below, determine whether or not functional B-gal will be produced in…
A: DNA is the genetic material in most living organisms. It is the information hub of the cell that…
Q: Using the plasmid map of pBCH2.0 provided above, predict how many DNA fragments would be formed if…
A: Restriction enzyme are specific DNA cutters and extremely useful while exploring molecular biology…
Q: A plasmid that is both ampicillin and tetracyclineresistant is cleaved with PstI, which cleaves…
A: The tetracycline resistance gene is present in the recombinant plasmid. To select the recombinant…
Q: What role did the transposon play in the transfer of resistance?
A:
Q: Primers designing for epitope tagging: Design forward and reverse primers to amplify the following…
A: Primers should generally have the following properties: 1. Length of 18-24 bases. 2. 40-60% G/C…
Q: Figure 1 above is showing the plasmid map of pUC18. Discuss how screening is carried out if this…
A: pUC18/19 is an artificial plasmid cloning vector. This plasmid vector comprises of Ampicillin [AmpR}…
Q: In E. coli, the gene bioD+ encodes an enzyme involved in biotin synthesis, and galK+ encodes an…
A: Gene transfer is the process of insertion of unrelated DNA into cells. There are three mechanisms of…
Q: You have another circular plasmid. Complete and effective digestion of this plasmid with a…
A: The restriction enzymes are endonuclease that means they can cut the DNA from the middle. They are…
Q: he Pfizer Covid-19 vaccine is one of the vaccines currently being rolled out for mass vaccination in…
A: Vaccine -- First human vaccine were used against virus was attenuated viruses .Vaccine can be…
Q: Predict the sizes of the plasmid cleavage products generated on digestion of pBR322 and pGLO with…
A: Plasmids are naturally occurring extra chromosomal DNA molecules and are mostly used for cloning…
Q: Explain the role of the gene that is the selectable marker gene in this experiment.
A: Introduction In the RDT (recombinant DNA technology), we insert the segment of DNA into the vector…
Q: The genotypes of five merozygote bacteria are shown below. In each set, the genotype of the lac…
A: In bacteria and their viruses, an operon is a genetic regulatory structure in which genes coding for…
Q: Can you give an example of a modified (tagged) primers to amplify E. coli UidA sequence and subclone…
A: Few important points to know . Which are as follows: Vector is DNA molecule that act as…
Q: If the band on the Northern blot for mRNA isolated from liver tissue is 2580 bp, whereas from brain…
A: Splicing The removal of intron, the non coding sequence of DNA present in the premature RNA before…
Q: Table 1 shows a list of restriction endonucleases with their recognition sequence and the sites of…
A: Restriction enzymes are those that cleave the double stranded DNA at specific site. They have…
Q: You first need to create a plasmid. What are the minimal components necessary to develop this…
A: Since you have asked many questions, I'm giving you the answer of first 3 Please repost in order to…
Q: The total size of the plasmid is 2686 bp. There is a PstI recognition site at position 439, HindIII…
A: Answer - d. 3 fragments: 447bp, 507bp and 1732bp
Q: Explain the process of how X-gal screening works with pUC19, you may build a model with boxes and…
A: Hello. Since your question has multiple parts, we will solve the first question for you. If you want…
Q: If an enhancer is placed on one plasmid and its corresponding promoter is placed on a second plasmid…
A: Plasmids are extrachromosomal DNA present in prokaryotic cells. Plasmids generally confer resistance…
Q: antibodies were produced in a rabbit against a 25 KDa human soluble protien. when the rabbit…
A: In western blotting, the target protein is recognized by a synthetic antibody, called a primary…
Q: Which of the following options best explains how the lacZ reporter gene works when screening for…
A: Given: Need to find the correct option among the given four options to know how lacZ reporter gene…
Q: By conducting conjugation experiments between Hfr and recipientstrains, Wollman and Jacob mapped the…
A: Horizontal gene transfer, also known as lateral gene transfer, is a process in which one bacterium…
Q: What size bands would you expect to see on the Southern blots using probe 1 and probe 2 for an…
A: Introduction The term Sickle cell disease defines the condition where due to a mutation, the blood…
Q: We are utilizing BL21 DE3 bacterial cells for the expression of the ADA protein via autoinduction.…
A: BL21 DE3 Bacterial cells: It is an E. coli B strain that lacks the Lon protease. Recombinant…
Q: You are cloning the genome of a new DNA virus into pUC18. You plate out your transformants on…
A: Introduction Plasmid Vectors: these are like the artificial vehicle to deliver our gene of insert…
Q: With the use of well-illustrated diagrams, reconstruct the entire cloning process by explaining…
A: Molecular cloning or cloning is a technique to assemble DNA of two different organisms and directing…
Q: You have set up a recombinant DNA experiment using the plasmid PBR322 as the vector (see plasmid…
A: PBR322 is a cloning vector which is widely used for cloning purposes. The map of PBR322 is given in…
Trending now
This is a popular solution!
Step by step
Solved in 3 steps
- When using a conventional plasmid cloning vector containing a b-galactosidase gene, it is possible to perform a "blue-white screen" to determine which bacteria have taken up a plasmid into which a DNA fragment as been inserted, as opposed to those that have taken up just reclosed plasmid vector, by growing the transformed cells on nutrient agar plates containing the artificial b-gal substrate X-gal. Will bacteria that have taken up a plasmid into which a DNA fragment has been inserted form a blue colony or a white colony when grown on this medium? Briefly explain why these bacteria would form a colony of the color you chose.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?With reference to the image below, discuss the process and principle involved for screening/selection of hosts (last stage of cloning) containing the intended recombinant plasmid. LacZ' = Gene for alpha-peptide of β-galactosidase.
- Explain the process of how X-gal screening works with pUC19, you may build a model with boxes and arrows. 2. You are utilizing BamHI (GGATCC) restriction site and HindIII (AAGCTT) restriction site. Within pUC19, BamHI is at position 263, while HindIII is at position 233. (Hint: position is like coordinate on a map). If you manage to insert GTF2H5 in pUC19 vector, what are the sizes of fragments if you digest the pUC19-GTF2H5 (this is after insertion) with the following restriction enzyme combination after gel electrophoresis: BamHI and HindIII There is an NdeI site right in the middle of the GTF2H5 that has been inserted in pUC19, what would be the fragment sizes, if you digest with BamHI and NdeI. HindIII and NdeI BamHI, HindIII, and NdeI. 3. Design an experiment to confirm the presence of insert GTF2H5 in pUC19 vector using the following method (besides restriction analysis above), assuming that you know the sequence of GTF2H5: Southern Blot Polymerase Chain ReactionOnce cloning is complete, a recombinant OXA-M290 protein can be produced in E. coli. Competent E. coli BL21(DE3) cells are transformed with the pET-28a-OXA-M290 plasmid, and the transformed cells are grown on an agar plate that contains kanamycin. The E. coli BL21(DE3) cells transformed with the pET-28a-OXA-M290 vector are next grown in liquid growth media that contains kanamycin. Once the culture reaches an OD600 of 0.6, isopropyl β-D-1-thiogalactopyranoside (IPTG) is added to a final concentration of 0.5 mM to induce protein production. How does the addition of IPTG result in the transcription of genes that are regulated by the T7 promoter in E. coli BL21(DE3) cells? [4 – 6 sentences suggested] For Context ONLY: For this biology course and based on the results obtained from a previous BLAST search, the bacteria responsible for the infection at a Hospital is most likely a new strain of Escherichia coli. This strain is given the name E. coli AFB. Earlier, BLAST was used to study a…Analyzing Cloned Sequences A base change (A to T) is the mutational event that created the mutant sickle cell anemia allele of beta globin. This mutation destroys an MstII restriction site normally present in the beta globin gene. This difference between the normal allele and the mutant allele can be detected with Southern blotting. Using a labeled beta globin gene as a probe, what differences would you expect to see for a Southern blot of the normal beta globin gene and the mutant sickle cell gene?
- 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?You receive purified mRNA encoding the peak protein of SARS Cov-2 as well as the plasmid pET32 (a +), how will you synthesize the cDNA of the peak protein and then clone in the pET32 (a +) vector
- With the use of well-illustrated diagrams, reconstruct the entire cloning process by explaining different stages of the cloning process that involves the following: d. Screening/selection of hosts containing the intended recombinant plasmid. For this stage(d), discuss the importance of a second marker that can be used for screening of genomic DNA for colonies containing the pka-1 under the principle of insertional inactivation. This should be properly explained using all the attributes of the plasmid described above.a) If lacZ was successfully inserted into the plasmid and E. coli successfully transformed, then the lacZ gene will be expressed which will breakdown X-gal producing a blue-colored colony. b) If the gene of interest was successfully inserted into the plasmid and E. coli successfully transformed, then the LacZ gene will not be expressed so X-gal will be broken down leading to a blue-colored colony. c) If the gene of interest was successfully inserted into the plasmid and E. coli successfully transformed, then the LacZ gene will not be expressed so X-gal will not be broken down leading to a white-colored colony. d) If lacZ was successfully inserted into the plasmid and E. coli successfully transformed, then the lacZ gene will be expressed which will breakdown X-gal producing a white-colored colony.antibodies were produced in a rabbit against a 25 KDa human soluble protien. when the rabbit antiserum was used in a western blot of human soluble protiens, it detected 25-KDa protien,but also bound to protiens of 65KDa and 39 KDa.assuming that a pure protien was used to make the antibodies,how do yuo explain the results of the western blot?