Biochemistry
Regulating Gene Expression
Compiled By Simon Mwangi
Edition: 2024/25
, Science | Biochemistry I of IV pages
1. In regards to the cDNA library problem, if one wants to express a eukaryotic protein in E. coli most
The genes contain introns.
note: the way around this problem is to make cDNA using reverse trasncriptase
2. The first step in creating the cDNA library is isolating the total ___ from a cell
mRNAs
3. In the second step of the cDNA library, the mRNAs are converted to cDNA by reverse transcriptase and
primer
4. This nicks the RNA, leaving segments as primer for DNA polymerase I
RNase H
5. This fills in the blanks, removes primers, and fills again
DNA pol I
6. This makes the DNA blunt ended
T4 poly
7. Adding linkers prepares the cDNA for ___ into a vector
cloning
8. How could one isolate a gene for a particular protein with an unknown nucleotide sequence?
synthesize an oligodeoxynucelotide probe which would be complimentary to one of the strands of the cloned
9. Which oligonucleotides should be used when making the probe?
sequences that contain AAs with the least degenerate codons (such Trp and Met; Asn and Lys have minimum
10. The Radioactive-antibody test. Describe process
a plastic disk coated with anti-insulin antibody is touched to the surface of colonies on a petri dish
insulin molecules from an insulin-producing colony bind to antibodies
radioactivity adheres to the disk at the positions of insulin-producing colonies
11. PCR permits as high as ___ fold increase in the concentration of a single DNA molecule
10^10
12. This PCR technique produces a fingerprint of an organism that would allow identification of disease
organisms. - With the set of short, random primers, different organisms will produce unique PCR products
RAPD analysis (random amplified polymorphic DNA)
13. In regards to use of PCR in genetic testing, ___ can be designed to detect specific ___ in affected patients
primers; mutations
14. PCR is so sensitive that only one or few copies of target DNA are required. True or false?
true
15. The DNA polymerase enzyme is isolated from ___ bacteria so that it is able to resist denaturation even in
Biochemistry 2024/25 Edition
Regulating Gene Expression
Compiled By Simon Mwangi
Edition: 2024/25
, Science | Biochemistry I of IV pages
1. In regards to the cDNA library problem, if one wants to express a eukaryotic protein in E. coli most
The genes contain introns.
note: the way around this problem is to make cDNA using reverse trasncriptase
2. The first step in creating the cDNA library is isolating the total ___ from a cell
mRNAs
3. In the second step of the cDNA library, the mRNAs are converted to cDNA by reverse transcriptase and
primer
4. This nicks the RNA, leaving segments as primer for DNA polymerase I
RNase H
5. This fills in the blanks, removes primers, and fills again
DNA pol I
6. This makes the DNA blunt ended
T4 poly
7. Adding linkers prepares the cDNA for ___ into a vector
cloning
8. How could one isolate a gene for a particular protein with an unknown nucleotide sequence?
synthesize an oligodeoxynucelotide probe which would be complimentary to one of the strands of the cloned
9. Which oligonucleotides should be used when making the probe?
sequences that contain AAs with the least degenerate codons (such Trp and Met; Asn and Lys have minimum
10. The Radioactive-antibody test. Describe process
a plastic disk coated with anti-insulin antibody is touched to the surface of colonies on a petri dish
insulin molecules from an insulin-producing colony bind to antibodies
radioactivity adheres to the disk at the positions of insulin-producing colonies
11. PCR permits as high as ___ fold increase in the concentration of a single DNA molecule
10^10
12. This PCR technique produces a fingerprint of an organism that would allow identification of disease
organisms. - With the set of short, random primers, different organisms will produce unique PCR products
RAPD analysis (random amplified polymorphic DNA)
13. In regards to use of PCR in genetic testing, ___ can be designed to detect specific ___ in affected patients
primers; mutations
14. PCR is so sensitive that only one or few copies of target DNA are required. True or false?
true
15. The DNA polymerase enzyme is isolated from ___ bacteria so that it is able to resist denaturation even in
Biochemistry 2024/25 Edition
