Cell Bio - Unit 2 - Dna Technology

Front 1

What is the source of homogenous cells for studying genes, proteins, molecular mechanisms, etc.

Back 1

Cell Cultures

Front 2

Hybridization on DNA microarrays locates _____ of ______ ______.

Back 2

Expression of multiple genes.

Front 3

What do restriction nucleases allow?

Back 3

Cleavage of DNA at specific sites.
*Bacterial enzymes that cut (hydrolyze) DNA at specific sequences of 4-8 nucleotides (restriction site)

Front 4

What is nucleic acid hybridization used for?

Back 4

Locate genes on DNA, using labeled DNA or RNA.
-based on complementary base
pairing.

Front 5

What is PCR used for?

Back 5

Cloning DNA.

Front 6

What is DNA engineering?

Back 6

Modification of DNA sequences by
insertion into cellular DNA.

Front 7

List properties of differentiated cells in a culture.

Back 7

• maintain properties characteristic for cell type
• have finite life time in culture
• can be “immortalized” and propagated for long time in culture.

Front 8

List properties of undifferentiated cells in a culture.

Back 8

• can divide and grow indefinitely in culture
• can be stimulated to differentiate to a different cell types

Front 9

What type of cells can be propagated indefinitely in culture?

Back 9

Malignant tumor cells.

Front 10

List the five restriction nucleases from the notes.

Back 10

Eco RI; Hind III; Not I; Alu 1; Hae III

Front 11

DNA restriction fragments visible by ___ ___________.

Back 11

Gel electrophoresis.

Front 12

What shows locations of restriction sites for one enzyme in relationship to restriction sites of other enzymes?

Back 12

Restriction map.

Front 13

What ultimately makes DNA technology possible?

Back 13

The existence of restriction nucleases.

Front 14

What do Dideoxyribonucleotides lack?

Back 14

3’ hydroxyl group.

Front 15

What is the most efficient way of determining a nucleotide sequence?

Back 15

Dideoxy (Enzymatic) Method
*Uses four different dideoxyribonucleotides (ddATP, ddGTP, ddCTP, ddTTP) to randomly terminate replication in vitro.

Front 16

Dideoxy method produces a collection of different DNA copies that terminate at every position in the original DNA (different in length by a single nucleotide). The DNA copies separated on the basis of their length using _______.

Back 16

PAGE - Polyacrylamide gel electrophoresis.

Front 17

Speed of robot DNA sequencers is limited by what?

Back 17

The size of the gel.
*Maximum = few hundred.

Front 18

What is the method used for locating genes within the genome (isolated or in situ)?

Back 18

Nucleic Acid Hybridization (Renaturation).

Front 19

What is the common laboratory procedure used to visualize the Nucleic Acid Hybridization?

Back 19

Southern blotting.

Front 20

What is "In situ hybridization" used for?

Back 20

It locates DNA base sequences on chromosomes, within cells and tissues, reveals the distribution of RNA in cells.

Front 21

List 2 probes.

Back 21

DNA & RNA.

Front 22

List 2 labels.

Back 22

Radioactive and Fluorescent.

Front 23

Hybridization on DNA micro arrays allows for the determination of:

Back 23

*which genes are being transcribed into mRNA
*which are silent
during cell differentiation, life and neoplastic transformation, and in response to different factors (growth factors, toxins, hormones, infection).

Front 24

What is cDNA?

Back 24

DNA complementary to mRNA extracted from cells.

Front 25

What is a cloning vector?

Back 25

A self-replicating genetic element, which can accept a foreign DNA fragment.

Front 26

What is transfection?

Back 26

The process of willingly introducing nucleic acids into cells.
EX. Recombinant DNA placed in bacterium.

Front 27

What are the differences in Genomic and cDNA libraries?

Back 27

cDNA only contains exons, but the Genomic DNA Library contains exons, introns, regulatory sequences, and spacer DNA.

Front 28

Define: locus.

Back 28

A locus (plural loci) is the specific location of a gene or DNA sequence on a chromosome.

Front 29

What is VNTR?

Back 29

VNTR (variable number of tandem repeat aka hypervariable microsatellite sequences): Common repeated DNA sequences.

Front 30

Which is the more accurate method for DNA sequencing: Shotgun sequencing or “clone-by-clone” approach?

Back 30

“Clone-by-clone” approach.

Front 31

What does BAC stand for?

Back 31

Bacterial artificial chromosomes.

Front 32

Define: Annotation.

Back 32

Annotation - process of interpreting a genome sequence by locating its genes and identifying their functions.

Front 33

T/F?
Functional DNA sequences are highly conserved.

Back 33

True.

Front 34

What are expression vectors?

Back 34

Generally a plasmid that is used to introduce a specific gene into a target cell.
*Can be used to transfect bacteria or eucaryotic cells.

Front 35

Fact: Insertion of artificially created DNA sequences (combination of naturally occurring fragments, natural + synthetic DNA, or totally synthetic) into the genomes of cells (somatic or germ cells) can produce rare proteins.
List some of the proteins created using this method.

Back 35

Clotting factor VIII protein, insulin, growth factors, viral coat proteins for vaccine production.

Front 36

What is a reporter gene?

Back 36

A gene whose activity can be monitored.

Front 37

What is green fluorescent protein (GFP) used for?

Back 37

It is used as a reporter protein, when determining the function of genes.
Ex. GFP joined to the promoter of gene in fruit fly that is active only in specialized set of neurons.

Front 38

What can site-directed mutagenesis reveal?

Back 38

• Functional properties of protein
• Protein-ligand interaction
• Importance of particular mutation for protein folding pattern
• Catalytic properties/sites

Front 39

Define: transgenic organisms.

Back 39

Organisms containing new genes or altered genes obtained through recombinant DNA techniques.

Front 40

What is the complete and permanent inactivation of a particular gene?

Back 40

Gene knockout.
*Knockout mice - very useful in research, especially identification of the role of genes in physiology.