In this lab, the DDX3 insert was to be shuttled from an expression vector we previously...
- In this lab, the DDX3 insert was to be shuttled from an expression vector we previously made into a different vector. How do you think we obtained the original human DDX3 insert? (5 points)
Homework Answers
DDX3 is a cofactor for HIV-1 replication. According to the given question, the DDX3 mutants are biochemically characterized by the helicase and nucleic acid requisite ATPase. The role of DDX3 and motif I provide a piece of confirmation for its implication of HIV-1 replication and nucleic acid-binding. The human DDX3 that lacks this domain will bind HIV-1 RNA with low affinity. Also, a specific ligand of peptide for the insertion that is selected by the phage display hinder with the HIV-1 replication after the transduction into the Hela P4 cells. Besides expansion that understanding of the structural function relationship of the protein, which results in identifying a particular domain of DDX3 that is suited as a target for the antiviral drugs designed to inhibit cellular co-factor for the HIV-1 replication.
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In this lab, the DDX3 insert was to be shuttled from an
expression vector we previously...
3. The pCMV-Ha vector and the DDX3 insert were cut with both EcoR1 and Sali, and...
3. The pCMV-Ha vector and the DDX3 insert were cut with both EcoR1 and Sali, and then purified. Why do you think we chose to cut with two different restriction enzymes, rather than just one (e.g. only EcoR1)? (7 points)
Now let's take a completely different approach to this problem. If we can find the vector...
Now let's take a completely different approach to this problem. If we can find the vector equation for the electric field at point P due to the ring of charge, then we ca can use the expression X --Cs AV V - V E-ds Jao as an alternative way to find a general equation for the potential at point P. da Activity 5.3: AV from a Ring using the E-field Method Show that the electric field at point P from...
8. (0.5 pts) One of the problems that we face is the option of the vector...
8. (0.5 pts) One of the problems that we face is the option of the vector closing on itself with no insert. We address that by changing ratios of insert and vector. What do we do and how does it help? 9. (0.5 pts) What are some other features of pJet1.2 plasmid vector that aid in making sure that the vector does not close on itself?
The molar ratio of insert DNA to vector DNA used in a ligation is generally about...
The molar ratio of insert DNA to vector DNA used in a ligation is generally about 3:1. Using a vector that is 12 kb and an insert that is 4 kb, how much insert would you use if you used 30 ng of vector? Give your answer in ng and only write the number, do not include the units.
PART 3 -SCREENING POTENTIAL CLONES (2 MARKS). After completing a cloning experiment, we screen potential clones...
PART 3 -SCREENING POTENTIAL CLONES (2 MARKS). After completing a cloning experiment, we screen potential clones to determine which ones are successful. The experiments outlined in Session 2-4 in the lab manual describe how we do this for the LEAPS plasmid. In this experiment using just the EcoRI site to facilitate cloning we have an additional problem - both ends of the PCR product have an EcoRI site and this means that the amplified sequence can be cloned into the...
Background In this lab, we are going to learn how to use a powerful separation technique...
Background In this lab, we are going to learn how to use a powerful separation technique called chromatography which can separate compounds that are quite similar to one another. Chromatography is a general type of separation in which a mixture of compounds passes through a stationary phase. Different compounds have different relative affinities for the mixture traveling along the stationary phase, or the mobile phase, and the stationary phase, causing these compounds to separate from one another. More specifically, we’re...
i honestly have no idea how to answer this one. can someone please help me out with a thorough answer so that i can lea...
i honestly have no idea how to answer this one. can someone
please help me out with a thorough answer so that i can learn from
it as well.
You have a forward primer that anneals within sticky fingers and a reverse primer that anneals within the Pen-tag (see pictures below). Xba1 EcoR1 Xho1 Sal1 Pst1 Tbd1 ReversePen-tag Forward primer 40 Sticky fingers e) We used blue white selection to figure out which colonies of E.col had our insert in...
Discussion questions Due at the beginning of next weeks' lab 1 How do expression vectors differ...
Discussion questions Due at the beginning of next weeks' lab 1 How do expression vectors differ from vectors used for replicating DNA? (1 mark) 2 What is a fusion protein expression vector? What is the fusion component of pET28a? (2 mark) 3 Briefly discuss some potential problems in expressing eukaryotic proteins in E. coll. (2 marks) 4 Briefly describe a His-tag and how it interacts with its target. (2 mark) 5 How does imidazole act to elute the His-tagged protein...
What units will we use for the forces in this lab? What is the meaning of...
What units will we use for the forces in this lab? What is the meaning of the Equilibrant? How do the Equilibrant and Resultant vectors relate to each other? What are the three approaches we will use to analyze the vector addition of forces in this lab? Describe them briefly.
We have been doing PCR to take DNA from Listeria monocytogenes and eventually add it to...
We have been doing PCR to take DNA from Listeria monocytogenes and eventually add it to e coli. We had a vector of pET28a and our PCR insert was HupD. Prior to purifying the PCR insert, it had a bp of 1700. We then purified our vector using a Qiagen mini-prep kit. We purified our PCR insert using a Qiagen PCR clean-up kit. We put both DNA solutions in the freezer until next class. In the next class, we added...
3. The pCMV-Ha vector and the DDX3 insert were cut with both EcoR1 and Sali, and then purified. Why do you think we chose to cut with two different restriction enzymes, rather than just one (e.g. only EcoR1)? (7 points)
Now let's take a completely different approach to this problem. If we can find the vector equation for the electric field at point P due to the ring of charge, then we ca can use the expression X --Cs AV V - V E-ds Jao as an alternative way to find a general equation for the potential at point P. da Activity 5.3: AV from a Ring using the E-field Method Show that the electric field at point P from...
8. (0.5 pts) One of the problems that we face is the option of the vector closing on itself with no insert. We address that by changing ratios of insert and vector. What do we do and how does it help? 9. (0.5 pts) What are some other features of pJet1.2 plasmid vector that aid in making sure that the vector does not close on itself?
PART 3 -SCREENING POTENTIAL CLONES (2 MARKS). After completing a cloning experiment, we screen potential clones to determine which ones are successful. The experiments outlined in Session 2-4 in the lab manual describe how we do this for the LEAPS plasmid. In this experiment using just the EcoRI site to facilitate cloning we have an additional problem - both ends of the PCR product have an EcoRI site and this means that the amplified sequence can be cloned into the...
i honestly have no idea how to answer this one. can someone
please help me out with a thorough answer so that i can learn from
it as well.
You have a forward primer that anneals within sticky fingers and a reverse primer that anneals within the Pen-tag (see pictures below). Xba1 EcoR1 Xho1 Sal1 Pst1 Tbd1 ReversePen-tag Forward primer 40 Sticky fingers e) We used blue white selection to figure out which colonies of E.col had our insert in...
Discussion questions Due at the beginning of next weeks' lab 1 How do expression vectors differ from vectors used for replicating DNA? (1 mark) 2 What is a fusion protein expression vector? What is the fusion component of pET28a? (2 mark) 3 Briefly discuss some potential problems in expressing eukaryotic proteins in E. coll. (2 marks) 4 Briefly describe a His-tag and how it interacts with its target. (2 mark) 5 How does imidazole act to elute the His-tagged protein...
What units will we use for the forces in this lab? What is the meaning of the Equilibrant? How do the Equilibrant and Resultant vectors relate to each other? What are the three approaches we will use to analyze the vector addition of forces in this lab? Describe them briefly.
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