Dr. Sharp- BioTech Exam 2
QUESTIONS N ANSWERS RATED A+
What makes a good vector? - correct answer ✔✔ 1. Size
2. Origin of replication (ori)
3. Multiple cloning sites
4. Selectable marker genes
5. RNA polymerase
6. Promotor sequences
7. DNA sequencing primer sequences
What is the origin of replication? - correct answer ✔✔ site for DNA replication
- allows plasmids to replicate independently from host chromosomes
What is a copy number? - correct answer ✔✔ number of plasmids per cell
- normally small
Describe multiple cloning site (MCS) - correct answer ✔✔ recognition site for several restriction
enzyme, in which DNA insert is cloned into
Describe selectable marker gene - correct answer ✔✔ are allow to select for transformed
colonies
Describe RNA polymerase promoter sequences (Rpps) - correct answer ✔✔ used for
transcription in vitro and in vivo
,Assume you want to make lots of human insulin using a bacteria expression vector. Why
wouldn't using human insulin genomic DNA for this clone project be advantageous? - correct
answer ✔✔ Introns and Exons
- cDNA comes from mRNA that corresponds to the human gene.
What prevents restriction enzymes produced by a bacteria from digesting its own genome?
A. Restriction preventase
B. Medichlorians
C. Acetylated histones
D. Methylated DNA
E. None of the above - correct answer ✔✔ D. Methylated DNA
What are the two types of DNA libraries? - correct answer ✔✔ 1. Genomic
2. Complementary DNA (cDNA) Libraries
What are DNA libraries? - correct answer ✔✔ collection of cloned DNA fragments
- from a particular organism contained in plasmid vectors within host bacteria
Ex: E. coli
What can DNA libraries be used for? - correct answer ✔✔ screen library to pick out different
gens of interest
Describe Genomic libraries - correct answer ✔✔ Chromosomal DNA from the tissue of interest
is isolated and digested with a restriction enzyme
- which produces many fragments that include the entire genome
What is used to ligate chromosomal DNA fragment within vector DNA (plasmids)?
, A. DNA ligase
B. DNA polmerase
C. Helicase
D. RNA primer - correct answer ✔✔ A. DNA ligase
What are used to transform bacteria, and theoretically each bacteria will contain a single
recombinant plasmid?
A. Protein
B. Recombinant vector
C. Copy number
D. None of the above - correct answer ✔✔ B. Recombinant vector
Disadvantages of genomic libraries - correct answer ✔✔ Introns & Exons are cloned
-Majority of genomic DNA is introns in eukaryotes so majority will have non coding pieces of
DNA
- Many organisms have a large genome
- It's time consuming
Describe cDNA libraries - correct answer ✔✔ mRNA from tissue of interest is extracted
- make double strand DNA from mRNA
How to make double stranded DNA from mRNA? - correct answer ✔✔ 1. Short linker double
stranded DNA sequences, which contain restriction enzyme recognition sites, are added to the
ends of the cDNA.
2. Cut with restriction enzyme, cut vector with same enzyme, ligate fragments to create
recombinant vectors
QUESTIONS N ANSWERS RATED A+
What makes a good vector? - correct answer ✔✔ 1. Size
2. Origin of replication (ori)
3. Multiple cloning sites
4. Selectable marker genes
5. RNA polymerase
6. Promotor sequences
7. DNA sequencing primer sequences
What is the origin of replication? - correct answer ✔✔ site for DNA replication
- allows plasmids to replicate independently from host chromosomes
What is a copy number? - correct answer ✔✔ number of plasmids per cell
- normally small
Describe multiple cloning site (MCS) - correct answer ✔✔ recognition site for several restriction
enzyme, in which DNA insert is cloned into
Describe selectable marker gene - correct answer ✔✔ are allow to select for transformed
colonies
Describe RNA polymerase promoter sequences (Rpps) - correct answer ✔✔ used for
transcription in vitro and in vivo
,Assume you want to make lots of human insulin using a bacteria expression vector. Why
wouldn't using human insulin genomic DNA for this clone project be advantageous? - correct
answer ✔✔ Introns and Exons
- cDNA comes from mRNA that corresponds to the human gene.
What prevents restriction enzymes produced by a bacteria from digesting its own genome?
A. Restriction preventase
B. Medichlorians
C. Acetylated histones
D. Methylated DNA
E. None of the above - correct answer ✔✔ D. Methylated DNA
What are the two types of DNA libraries? - correct answer ✔✔ 1. Genomic
2. Complementary DNA (cDNA) Libraries
What are DNA libraries? - correct answer ✔✔ collection of cloned DNA fragments
- from a particular organism contained in plasmid vectors within host bacteria
Ex: E. coli
What can DNA libraries be used for? - correct answer ✔✔ screen library to pick out different
gens of interest
Describe Genomic libraries - correct answer ✔✔ Chromosomal DNA from the tissue of interest
is isolated and digested with a restriction enzyme
- which produces many fragments that include the entire genome
What is used to ligate chromosomal DNA fragment within vector DNA (plasmids)?
, A. DNA ligase
B. DNA polmerase
C. Helicase
D. RNA primer - correct answer ✔✔ A. DNA ligase
What are used to transform bacteria, and theoretically each bacteria will contain a single
recombinant plasmid?
A. Protein
B. Recombinant vector
C. Copy number
D. None of the above - correct answer ✔✔ B. Recombinant vector
Disadvantages of genomic libraries - correct answer ✔✔ Introns & Exons are cloned
-Majority of genomic DNA is introns in eukaryotes so majority will have non coding pieces of
DNA
- Many organisms have a large genome
- It's time consuming
Describe cDNA libraries - correct answer ✔✔ mRNA from tissue of interest is extracted
- make double strand DNA from mRNA
How to make double stranded DNA from mRNA? - correct answer ✔✔ 1. Short linker double
stranded DNA sequences, which contain restriction enzyme recognition sites, are added to the
ends of the cDNA.
2. Cut with restriction enzyme, cut vector with same enzyme, ligate fragments to create
recombinant vectors
