In this western blot, the levels of TBK remain constant with increasing amounts/expression 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.VPPTBK1(S172)TBK1actinVirusproteinsO TrueO False

TBK-1 gene encodes an enzyme known as TBK-1 (TANK binding kinase-1). This is a…

Answered: In this western blot, the levels of TBK…

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

Chapter17: Bacterial And Viral Genetics

Section: Chapter Questions

Problem 1ITD

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Below is a map of the E. coli cloning vector after the insertion of pka-1. Ori denotes the origin of replication; amp denotes the ampicillin resistance gene. HindIII, BamHI, SalI, and EcoRI designate restriction enzyme sites. There are no other restriction enzyme sites found on this vector. The numerals denote the number of base pairs between different locations on the plasmid. For instance, there are 400 base pairs between the HindIII and BamHI site, and there are 3000 base pairs in the entire cloning vector (following the integration of pka-1). With the use of well-illustrated diagrams, reconstruct the entire cloning process by explaining different stages of the cloning process that involves the following: Isolation of target DNA fragments (often referred to as inserts) Ligation of inserts into the plasmid, creating recombinant molecules Transformation of recombinant plasmids into bacteria or other suitable host for propagation Screening/selection of hosts containing the intended…
As a part of undergrad lab project, you have to clone Your Favorite Gene into plasmid cloning vector (4740 bp) pUC18. Restriction maps for both YFG and cloning vector are provided. The plasmid has a bacterial origin of replication (ori), an ampicillin resistance gene (AmpR) that degrades ampicillin antibiotic. It also has LacZ gene that encodes the β-gal enzyme, which converts white X-gal substrate into a blue product. pLac is a bacterial promoter. Restriction Enzyme Recognition Site : A slash (/) represents the cut site for each restriction enzyme. Enzyme 1 5’- G/TCGA C -3’ 3’- C AGCT/G-5’ Enzyme 2 5’-C/TCGA G-3’ 3’G AGCT/ C5’ Enzyme 3 5’- GTC / GAC- 3’ 3’- CAG / CTG- 5’ Enzyme 4 5’-C AATT /G-3’ 3’-G/ TTAA C-5’ Enzyme 5 5’C AATT/G-3’ 3’G/TTAA C-5’ Enzyme 6 5’G/GATC C-3’ 3’C CTAG/ G-5’ Enzyme 7 5’G/GATC C-3’ 3’C CTAG/ G-5’ 1a) The table below shows restriction enzyme pairs used by various teams to digest the YFG and clone it in the…
A Western blot is seen in the picture above. Sodium dodecyl sulfate gel polyacrylamide gel electrophoresis was used to isolate the proteins (SDS-PAGE).1. Since gel electrophoresis has isolated individual proteins, what kind of molecule is used to detect them? 2. On the right side of this Western blot, three molecules are identified. One of the bands corresponds to the smallest molecule?
When cloning a foreign DNA fragment into a plasmid, it is often useful to insert the fragment at a site that interrupts a selectable marker(such as the tetracycline-resistance gene of pBR322). The loss of function of the interrupted gene can be used to identify clones containing recombinant plasmids with foreign DNA. With a yeast artificial chromosome (YAC) vector, it is not necessary to do this; the researcher can still distinguish vectors that incorporate large foreign DNA fragments from those that do not. How are these recombinant vectors identified?
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 Reaction
A student cloned Claudin 1 (877 bp) gene into pTRE3G-BI-ZsGreen1 (total size is 8905 bp) vector using Not1 and EcoRV restriction sites located at 215bp and 779bp respectively. What is the size of the recombinant plasmid? A) 9782 bp B) 9218 bp C) 10776 bp E) 8028 bp F) None of the answers given
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?
You receive purified mRNA that codes for the spike protein of SARS-CoV-2 and the plasmid pET32(a+), how will you synthesize and clobe the cDNA of the spike protein into the pET32(a+) vector.
The map of plasmid pUC19 is shown below. The restriction site coordinate is the position of the 5’base on the top strand of each site sequence. The restriction enzyme sites are in bold type if there is only one site in pUC19. Please list the fragments in order of size, largest to smallest, which will result from a complete digestion by the restriction enzyme PvuII. Please list the fragments in order of size, largest to smallest, which will result from a complete digestion by the restriction enzyme DrdI.
The total size of the plasmid is 2686 bp. There is a PstI recognition site at position 439, HindIII recognition site at position 447, and ScaI recognition site at 2179. If restriction enzymes ScaI and HindIII are used to cut this plasmid, what would be produced? a. 2 fragments: 954bp and 1732bp b. none of the choices apply c. 1 linear fragment of 2686bp d. 3 fragments: 447bp, 507bp and 1732bp
In E. coli, the genes for histidine (his), arginine (arg), tyrosine (tyr), and valine (val) synthesis are closely linked. A wild-type strain is used as a donor for transducing viruses. Various recipient strains were treated with these viruses. Wild-type recombinant colonies were then counted, with the results shown in the following table. Genotype of recipient Number of wild-type colonies Genotype of recipient Number of wild-type colonies his- arg+ val+ tyr+ 254 his+ arg- val- tyr+ 132 his+ arg+ val- tyr- 36 his+ arg- val+ tyr+ 240 his- arg- val+ tyr+ 28 his- arg+ val- tyr+ 102 his- arg+ val+ tyr- 100 his+ arg- val+ tyr- 2 What is the cotransduction coefficient of histidine and valine? For consistency, standardize on histidine. 0.38 (wrong) 0.537 0.402 None of these
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?

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