You are asked to create a vector that will express the protein called DCoH in bacteria ( https://www.ncbi.nlm.nih.gov/nuccore/50086629 ).
Target: use pET28b ( https://www.staff.ncl.ac.uk/p.dean/pET28.pdf ) construct the vector so that there are no N-terminal additions and the DCoH protein will have a C-terminal Histidine tag.
Be sure to CLEARLY identify:
a) the sequence of your primers
b) the melting temperature of each of your primers
c) the restriction enzymes you plan to use with the sequences underlined in your primers
d) will your construct result in the production of C-terminally Histidine-tagged DCoH? prove it by in silico translating the last 2 codons of DCoH along with the rest of your tag
You are asked to create a vector that will express the protein called DCoH in bacteria ( https://www.ncbi.nlm.nih.gov/nuccore/50086629 ).
Target: use pET28b ( https://www.staff.ncl.ac.uk/p.dean/pET28.pdf ) construct the vector so that there are no N-terminal additions and the DCoH protein will have a C-terminal Histidine tag.
Be sure to CLEARLY identify:
a) the sequence of your primers
b) the melting temperature of each of your primers
c) the restriction enzymes you plan to use with the sequences underlined in your primers
d) will your construct result in the production of C-terminally Histidine-tagged DCoH? prove it by in silico translating the last 2 codons of DCoH along with the rest of your tag
For unlimited access to Homework Help, a Homework+ subscription is required.
Related textbook solutions
Related questions
Which of the following did Erwin Chargaff observe?
| ||
| ||
| ||
| ||
|
The melting temperature (Tm) of a DNA duplex is:
| ||
| ||
| ||
| ||
|
Given your knowledge and the descriptions of each of thesemethods, which of the following does NOT rely on the ability ofnucleic acids to hybridize with each other?
| ||
| ||
| ||
| ||
|
Which of the following is a possible reason why DNA uses thymineinstead of uracil?
| ||
| ||
| ||
|
Which of the following is an enzyme used in cloning to breakcovalent bonds?
| ||
| ||
| ||
| ||
|
Which of the following could you use to remove the 5' phosphatesafter cleavage of a plasmid with a single type of restrictionenzyme to ensure that the plasmid would not simply fuse backtogether upon addition of ligase?
| ||
| ||
| ||
| ||
|
Which of the following would you use to create a cDNA librarythat you would not use for a genomic library?
| ||
| ||
| ||
| ||
|
A plasmid vector typically has which of the followingfeatures?
| ||
| ||
| ||
| ||
|
Which of the following is FALSE of genomic or cDNAlibraries?
| ||
| ||
| ||
| ||
|
Which of the following is NOT a typical component of apolymerase chain reaction (PCR)?
| ||
| ||
| ||
| ||
|
How can a fusion protein be formed?
| ||
| ||
| ||
| ||
|
Why is green fluorescent protein (GFP) so useful in visualizingfusion proteins in eukaryotes?
| ||
| ||
| ||
| ||
|
You wish to determine the cellular location of a protein butunfortunately it is not particularly antigenic (can t develop astrong antibody response). How could you solve this problem?
| ||
| ||
| ||
| ||
| ||
|
A positive result for the yeast two-hybrid analysis means thefollowing:
| ||
| ||
| ||
| ||
|
Which of the following is the approximate size of the humangenome?
| ||
| ||
| ||
| ||
|
Which of the following is correct about the structure orlocation of genes?
Question 22 options:
| ||
| ||
| ||
| ||
|