Biotech Test 1 Set 2

Front 1

Sickle Cell Anemia
a)type of disease
b)follows a...

Back 1

a)genetic, chronic and hemolytic disease
b)recessive trait, but both parents must be carriers of gene

Front 2

Electrophoresis result of Haemoglobin S when compared to normal hemoglobin and meaning?

Back 2

moves more slowly towards (+) anode than does normal hemoglobin; HbS is less (-) than normal hemoglobin

Front 3

Sicle Cell Carriers results on gel electrophoresis?

Back 3

2 bands b/c they have HbA and HbS

Front 4

Sickle Cell Hemoglobin diff from regular hemoglobin (2) and who discovered it?

Back 4

Valine where a Glutamate should be in the Beta chain; neutral AA for acidic AA

Vernon Igram

Front 5

Problems with Sickle Hemoglobin? (2)

Back 5

1)valine gives sticky patches on outside of each Beta chain
2)causes molecules of HbS to stack via H-bonds and distort the red cells

Front 6

Sickle Shape leads to... (6)

Back 6

1)rapid destruction of RBC's
2)shortage of breath
3)repeated infections
4)clumping of RBC's
5)enlarged heart/spleen
6)thrombosis

Front 7

____ is the most common lethal inherited genetic disease in whites

Back 7

cystic fibrosis

Front 8

Parents have to be what to give their kid CF?

Back 8

have to be carriers of the defective gene that causes CF

Front 9

What happens in lung when you have CF?

Back 9

cilia of lung unable to move mucus towards mouth

Front 10

Clinical symptoms of CF (6)

Back 10

1)progressive lung disease
2)chronic lung infections
3)GI obstruction
4)ineffective reproductive system
5)Lungs NORMAL @ birth
6)chronic cough

Front 11

Type of lung infections in CF patients and what is special about one of em? (3)

Back 11

1)staphylcoccus
2)streptococcus
3)pseudomonas aeroginosa (NOT TREATABLE WITH ABX)

Front 12

Progressive lung disease in CF patients steps (3)

Back 12

1)lungs normal @ birth
2)bronchial mucus block small airways and become inflamed
3)mucus stays in lungs and provides spot for bacterial infections which lead to lung damage

Front 13

95% of CF patients die of...

Back 13

respiratory failure

Front 14

Pancreatic Insufficiency steps in a CF? (4)

Back 14

1)mucus blocks pancreatic ducts
2)this blocks pancreatic enzymes from getting to SI
3)leads to malabsorption of nutrients
4)"failure to thrive"

Front 15

GI obstruction in CF steps (3)

Back 15

1)Meconiumileus (blockage of illium)
2)dark green substance that emerges as first stool becomes too thick and blocks intestine
3)requires surgery to remove obstruction

Front 16

Effect of CF on male reproductive system

Back 16

obstruction of genital tract makes vas deferens, epididymis, seminal vesicle atrophic/fibrotic

Front 17

Effect of CF on female reproductive system

Back 17

mucus plugs cervix which doesn't allow sperm to penetrate uterus

Front 18

Sweat test of CF (or IF FAMILY MEMBER HAS CF)
a)how test is done
b)what looking for

Back 18

a)put pilocarpine on skin and absorb sweat in piece of filter paper
b)measure amount of Na and Cl in sweat (CF patients have 4x more than normal)

Front 19

Properties of protein that is product of defective CF gene? (1)

Back 19

1)cAMP induced chloride channel

Front 20

Name of gene responsible for CF? and fxn

Back 20

cystic fibrosis transmembrane conductance regulator (CFTR)

chloride channel protein in membrane

Front 21

MAIN mutation responsible for CF

Back 21

deletion of Phe @ codon 508

Front 22

What does the deletion of Phe 508 at do to CFTR? (3)

Back 22

1)pivotal b/c Phe is hydrophobic
2)leads to conformation change in CFTR
3)so it is degraded in ribosomes

Front 23

Effect on cell of not having CFTR?

Back 23

Cl- remains in cell so more Na+ in cell to equalize cell b/c of the opposite charges

Front 24

_____ increases the melting point of DNA

Back 24

having more C-G pairs

Front 25

Nucleotides in DNA are linked by....

Back 25

phosphodiesters bonds

Front 26

Purines (2)

Back 26

A and G

Front 27

Pyrimidines (3)

Back 27

C,U,T

Front 28

Nucleotide consists of.... (3)

Back 28

Base
Pentose(sugar)
Phosphate

Front 29

Bonds in a nucleotide (2)

Back 29

n-glycoside bond links base and pentose sugar

phosphoester bond links phosphate and pentose sugar

Front 30

Basic mechanism of recombinant DNA technology

Back 30

transfer of DNA from one organism to another

Front 31

Step 1 of rDNA (2)

Back 31

1)DNA from donor is cut out via restriction endonuclease
2)joined via ligases to recipient DNA to form recombinant DNA

Front 32

Step 2 of rDNA (2)

Back 32

1)recombinant DNA transferred to a host
2)intro of rDNA to bacterial host plasmid is called transformation

Front 33

Step 3 of rDNA (2)

Back 33

1)host cell are identified and selected
2)selected hosts are then mass cultured by fermentation methods

Front 34

Step 4 of rDNA (1)

Back 34

If reqd, a DNA construct can be prepared to ensure that the protein product that is encoded by the cloned DNA sequence is produced by the host

Front 35

Step 5/6 of rDNA (2 total)

Back 35

1)isolate, purify and modify protein drug
2)formulate product, study stability, and establish optimal delivery method

Front 36

Hybridization based technologies (5)

Back 36

1)PCR
2)southern blot
3)northern blot
4)microarrays
5)siRNA (RNA silencing)

Front 37

Tm def. and is directly related to...

Back 37

temp. that causes DNA to separate by 50%

composition of DNA sequence

Front 38

Decent formula for determining Tm

Back 38

2(# of A-T pairs) + 4(# of G-C pairs)

Front 39

DNA and DNA-RNA hybrids are destabilized by....(4)

Back 39

1)decr salts concentrations
2)incr detergent concentrations
3)incr temperatures
4)sequence mismatches

Front 40

Then why use a 20 base primer for PCR?

Back 40

increases specificity and gives it a higher Tm than if using a smaller primer

Front 41

Type of DNA polymerase used in PCR and why?

Back 41

Taq DNA Polymerase b/c it is thermally stable

Front 42

Steps in PCR (5)

Back 42

1)Heat @ 94C to separate strands
2)Add synthetic primers and cool to 45-60C
3)Add thermostable DNA polymerase to catalyze 5'->3' DNA synthesis @ 72C
4)keep repeating these steps
5)72C for 5 minutes for completion of parital products

Front 43

# of copies of amplified DNA in PCR can be determined by this equation....

Back 43

(2^N)(y)

N= # of cycles
y=starting # of DNA templates

Front 44

Southern Blot
a)use
b)requires...(2)
b)who invented it

Back 44

a)detect presence of a specific sequence
b)"labeled" probe to detect target sequence
b1)probe can either be labeled radioactively or chemically
c)Dr. EM Southern

Front 45

Gel electrophoresis def.

Back 45

movement of an electrically charged substance under the influence of an electric field

Front 46

Gel electrophoresis steps for DNA (4)

Back 46

1)restriction enzymes cleave DNA into smaller pieces
2)DNA loaded into wells in a porous gel in a buffer solution
3)electric current activated and DNA moves towards (+) cathode
4)smaller DNA frags move faster and v.v.

Front 47

Northern blot difference from Southern blot (4)

Back 47

1)USES RNA AS SOURCE OF NUCLEIC ACID
2)RNA is not enzymatically digested
3)gel used to separated RNA molecules has denaturing agents in it so that post-treatment of gel is unnecessary
4)No Dr. Northern

Front 48

Microarrays
a)def.
b)what is a microarray
c)reverses....

Back 48

a)hybridization analysis of mRNA (or DNA) on a genome-wide scale
b)membrane, glass slide or "chip" containing probes for every gene predicted in a genome
c)reverses the hybridization - put the probes on the filter and label the total RNA

Front 49

____ does photolithograph

Back 49

GeneChip

Front 50

Sample isolation and labeling for Microarray (2)

Back 50

1)mRNA is isolated and converted to cDNA via reverse transcriptase
2)cDNA is then fluorescently labeled to allow detection on the array

Front 51

Microarray Applications (5)

Back 51

1)genome wide transcriptional profiling
2)comparison of mRNA from different tissues
3)comparison of normal and disease states
4)drug or environmental response studies
5)"low-level" genome sequencing

Front 52

Genetic source material for PCR

Back 52

DNA
cDNA

Front 53

Genetic source material for DNA Sequencing

Back 53

DNA
cDNA

Front 54

Genetic source material for Nucleic Acid Hydridization (north/south blots)

Back 54

DNA
RNA
cDNA NOT usually used but can be

Front 55

Genetic source material for Microarray (and name of analysis when diff material is used)

Back 55

DNA (genomic analysis)
cDNA (transcriptional analysis)