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33 Cards in this Set

  • Front
  • Back
Southern Blots detect what?
DNA sequence of interest
Western Blots detect what?
Protein of interest
Northern Blot detects what?
RNA of interest
PCR does what?
Amplifies DNA of interest
Define Cloning Vectors
Self-replicating unit of DNA that can be manipulated to incorporate a foreign DNA insert
List 3 types of cloning vectors that are used.
What are the requirements for cloning vectors? x3
Restriction site

Ability to get into cell

Selectable phenotype
What are the steps in Gene Cloning?
1. Choose DNA source

2. Produce and Propagate fragments

3. Screen for Desired Results
In terms of gene cloning, what type of DNA source would be valuable? x2
Chromosomal DNA

In terms of gene cloning, what steps would have to be taken to produce and propagate DNA fragments? x4
1. Restriction enzymes/reverse transcriptase

2. Insert fragments into vectors

3. Transform bacteria

4. Plate bacteria to antibiotics
What are the two ways to screen for desired results?
Direct: Nucleic Acid Probe
(based on screening library)

Indirect: Antibiotics to Protein
(based on protein produced)
What are the four approaches to Cancer Gene Therapy?
Suicide Selection

Induce genes encoding tumor suppressor genes (p53)

Introduce DNA to lower levels of activated oncogenes

Administer DNA to increase immune system to kill tumors.
What is the suicide selection approach?
A gene encoding an enzyme that can convert a compound into a toxin that can be used on tumor cells.
Benefits of the following?

- Northern blot
- Southern blow job
- Western blot
Detection and characterization of:

- Proteins
Benefits of the following?

- Branched chain PCR
Amplification of:

What Molecular techniques serve the purpose of DNA comparison? x2

DNA electrophoresis
In situ Hybridization serves what purpose?
Detection of Nucleic acids
Dot Blot serves what purpose?
DNA detection
SDS-PAGE/Western Blot serves what purpose?
Separation & Identification
Why clone DNA? x4
1. OBTAIN Fragment of DNA for PROBE

2. OBTAIN a DNA for Mutagenesis Study

3. PRODUCE a purified protein

Applications of Recombinant DNA. x4
1. Protein production
(insulin or G-CSF)

2. Understand Genes better
(structure, f(x), & regulation)

3. Rapid ID of microorganisms

4. Dx & Tx of Human Dz
What type of gene therapy is the most common area being explored now?
Cancer gene therapy
DNA microarrays has ALLOWED for the expression of????
Expression of 1000's of genes at one time.

Able to compare normal cells CA cells to determine changes in expression.
Microarrays provide what major benefit especially in CA? x2
1. Allows comparisons of normal genes with diseases ones (e.g. - CA)

2. Prognostic indicator.
(spares many pts from unnecessary chemo and radiation.
Give an example of a novel therapeutic that has developed as a result of Recombinant DNA technology assisting the understanding of Gene structure, f(x), and regulation.

(for the treatment of CML)
Glivec can be used to Tx?

(Chronic Myelogenous Leukemia)
Discuss what the colors mean in microarrays.
Green is decreased gene expression of CA cells

Red is increased gene expression in CA cells

Yellow indicates NO change
In a microarray, each spot represents what?
Different gene
What 3 questions must be satisfied in developing Gene Therapy?
1. Delivered to target cells AND Remain there long enough to be effective?

2. Expressed at High enough levels?

3. Harmful to patient?
Gene Therapy GENES can be introduced in what 2 ways?
1. Into cells recovered from patient (ex vivo)


2. In vivo into patient directly
What vectors are commonly used to deliver genes in Gene Therapy?
Viral vectors

(Liposomes & Naked plasmids are currently being attempted)
How many types of plasmids are there?
Over 1000
How many CA Gene Therapy trials are underway?
Over 100