Exam 3

Question 1

Restriction Endonucleases (enzymes)

Answer 1

• origination in bacterial cells
• Protect organism (bacteria) from foreign DNA (bacteriophage)
• Cleaves DNA at phospate-sugar bonds
• Recognizes palindromes of 4-10 base pairs where it cleaves DNA

Question 2

Sticky Ends

Answer 2

Unpaired bases that are left where restriction endonucleas cleaved at DNA palindrome

Question 3

Ligase

Answer 3

DNA ligase, rejoins base pairs

Question 4

Reverse Transcriptase

Answer 4

Converts RNA to DNA

Question 5

Four DNA analysis techniques

Answer 5

• electrophoresis
• hybridization and probes
• Sequencing
• PCR

Question 6

Electrophorsis

Answer 6

• DNA is cut with restriction endonucleauses to specific gene sequences
• put in gel at (-) end and phosphates (base pairs) move towards the (+) end
• Smaller sequences move farther

Question 7

Probes

Answer 7

• Small sequences of single stranded nucleic acids (olionucleotides)
• Used to identify complementary sequences on single stranded DNA

Question 8

Southern Blot

Answer 8

• Method for detecting unknown DNA sequences

- electrophesis seperates DNA, denatured and transfered to filter paper, probes to visualize hybridization

Question 9

DNA/RNA hybridization

Answer 9

-Degree of similarity between two different strands

Question 10

Sequencing

Answer 10

Identify and order nucleotides

-Sanger Method

Question 11

Polymerase Chain Reaction

Answer 11

• amplifiy’s DNA

- Can be used to detect diseases/infectious agents

Question 12

Recombinant DNA

Answer 12

-DNA genetically manufactured to possess genetic material from multiple sources

Question 13

Cloning Vectors

Answer 13

• Carry significant piece of donor DNA readily accepted by cloning host
• ex. plasmids, phages

Question 14

Cloning Host

Answer 14

Bacteria (prokaryote)
-E. coli
-will NOT excess introns from eukaryotic DNA
Yeast
-S. cerevisae
-will excess introns

Question 15

Gene Therapy

Answer 15

Used to repair genetic defects such as: immunodeficiency diseases, cycstic fibrosis, Sickle Cell Anemia

Question 16

In vivo Strategy

Answer 16

• Gene therapy technique

- Done inside a live organism (rat, gerbil, human, etc.)

Question 17

Ex vivo strategy

Answer 17

• Gene Theraoy technique

- Done with living cells but not in an organism

Question 18

Antisense DNA/RNA

Answer 18

• Gene therapy technique

- A oligonucleotide (single stranded DNA/RNA) that binds to mRNA to prevent specific protein synthesis

Question 19

Triplex DNA

Answer 19

-Targets double stranded DNA and prevents transcription

Question 20

Three types of Genome Analysis

Answer 20

Maps
fingerprints
family trees

Question 21

3 types of Genome Maps

Answer 21

• Linkage - relative proximity and locus of genes
• Physical- proximity and size of genes
• Sequence - exact order of bases

Question 22

Genome Maps

Answer 22

-Determine locus (location) and alleles

Question 23

Fingerprinting

Answer 23

Emphasizes differences in genome
-techniques
endonucleases
PCR
Southern Blot
-Uses
forensics
hereditary diseases

Question 24

Family Trees

Answer 24

• Determine genetic tree for diseases
• Animal pedigrees
• genetic diversity in animals

Question 25

Ideal Antimicrobic

Answer 25

``` -Soluble in body fluids selectively toxic nonallergenic reasonable half life unlikely to elecit resistance long shelf life good priced. ```

Question 26

Chemotherapy

Answer 26

Use of any chemical agent for disease treatment

Question 27

antibiotic

Answer 27

chemical substance made by microbe

Question 28

antimicrobic

Answer 28

synthetic version of a antibiotic

Question 29

Sources of antibiotics (3)

Answer 29

- mold type fungi (Penicillium, Cephalosporium) - Bacillus strains (bacitracin) - Actinomycetes (soil bacteria)

Question 30

Broad Spectrum Antibiotic

Answer 30

treatment is effect against wide variety of microbes

Question 31

Narrow Spectrum Antibiotic

Answer 31

Treatment is effective against a limited number of similar microbes

Question 32

Kirby-Bauer Disk Diffusion Test

Answer 32

-Antibiotic placed on plate with lawn of targeted bacteria, clear zones around antibiotic indicate effectiveness

Question 33

Minimum Inhibitory Concentration

Answer 33

-lowest concentration that will inhibit growth

Question 34

5 Mechanisms of Action for Antibiotics

Answer 34

- Inhibition of cell wall synthesis - disruption of cell membrane function - inhibition of nucleic acid synthesis - anti-metabolic activity

Question 35

Cell Wall antibiotics

Answer 35

- Penicillin, Cephalosporins - block transpeptididation (required in cell walls to cross link peptidoglycan) - Bacitracin utilizes different method due to topical application only

Question 36

Penicillins

Answer 36

- 1945 (1st drug) - Sir Alexander Flemming - beta-lactam ring w/ variable R group - Targets transpeptidation - Also triggers autolysin (digests cell wall)