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55 Cards in this Set
- Front
- Back
Sickle Cell Anemia
a)type of disease b)follows a... |
a)genetic, chronic and hemolytic disease
b)recessive trait, but both parents must be carriers of gene |
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Electrophoresis result of Haemoglobin S when compared to normal hemoglobin and meaning?
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moves more slowly towards (+) anode than does normal hemoglobin; HbS is less (-) than normal hemoglobin
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Sicle Cell Carriers results on gel electrophoresis?
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2 bands b/c they have HbA and HbS
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Sickle Cell Hemoglobin diff from regular hemoglobin (2) and who discovered it?
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Valine where a Glutamate should be in the Beta chain; neutral AA for acidic AA
Vernon Igram |
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Problems with Sickle Hemoglobin? (2)
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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 |
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Sickle Shape leads to... (6)
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1)rapid destruction of RBC's
2)shortage of breath 3)repeated infections 4)clumping of RBC's 5)enlarged heart/spleen 6)thrombosis |
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____ is the most common lethal inherited genetic disease in whites
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cystic fibrosis
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Parents have to be what to give their kid CF?
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have to be carriers of the defective gene that causes CF
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What happens in lung when you have CF?
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cilia of lung unable to move mucus towards mouth
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Clinical symptoms of CF (6)
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1)progressive lung disease
2)chronic lung infections 3)GI obstruction 4)ineffective reproductive system 5)Lungs NORMAL @ birth 6)chronic cough |
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Type of lung infections in CF patients and what is special about one of em? (3)
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1)staphylcoccus
2)streptococcus 3)pseudomonas aeroginosa (NOT TREATABLE WITH ABX) |
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Progressive lung disease in CF patients steps (3)
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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 |
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95% of CF patients die of...
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respiratory failure
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Pancreatic Insufficiency steps in a CF? (4)
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1)mucus blocks pancreatic ducts
2)this blocks pancreatic enzymes from getting to SI 3)leads to malabsorption of nutrients 4)"failure to thrive" |
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GI obstruction in CF steps (3)
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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 |
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Effect of CF on male reproductive system
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obstruction of genital tract makes vas deferens, epididymis, seminal vesicle atrophic/fibrotic
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Effect of CF on female reproductive system
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mucus plugs cervix which doesn't allow sperm to penetrate uterus
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Sweat test of CF (or IF FAMILY MEMBER HAS CF)
a)how test is done b)what looking for |
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) |
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Properties of protein that is product of defective CF gene? (1)
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1)cAMP induced chloride channel
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Name of gene responsible for CF? and fxn
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cystic fibrosis transmembrane conductance regulator (CFTR)
chloride channel protein in membrane |
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MAIN mutation responsible for CF
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deletion of Phe @ codon 508
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What does the deletion of Phe 508 at do to CFTR? (3)
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1)pivotal b/c Phe is hydrophobic
2)leads to conformation change in CFTR 3)so it is degraded in ribosomes |
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Effect on cell of not having CFTR?
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Cl- remains in cell so more Na+ in cell to equalize cell b/c of the opposite charges
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_____ increases the melting point of DNA
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having more C-G pairs
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Nucleotides in DNA are linked by....
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phosphodiesters bonds
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Purines (2)
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A and G
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Pyrimidines (3)
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C,U,T
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Nucleotide consists of.... (3)
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Base
Pentose(sugar) Phosphate |
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Bonds in a nucleotide (2)
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n-glycoside bond links base and pentose sugar
phosphoester bond links phosphate and pentose sugar |
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Basic mechanism of recombinant DNA technology
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transfer of DNA from one organism to another
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Step 1 of rDNA (2)
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1)DNA from donor is cut out via restriction endonuclease
2)joined via ligases to recipient DNA to form recombinant DNA |
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Step 2 of rDNA (2)
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1)recombinant DNA transferred to a host
2)intro of rDNA to bacterial host plasmid is called transformation |
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Step 3 of rDNA (2)
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1)host cell are identified and selected
2)selected hosts are then mass cultured by fermentation methods |
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Step 4 of rDNA (1)
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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
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Step 5/6 of rDNA (2 total)
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1)isolate, purify and modify protein drug
2)formulate product, study stability, and establish optimal delivery method |
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Hybridization based technologies (5)
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1)PCR
2)southern blot 3)northern blot 4)microarrays 5)siRNA (RNA silencing) |
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Tm def. and is directly related to...
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temp. that causes DNA to separate by 50%
composition of DNA sequence |
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Decent formula for determining Tm
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2(# of A-T pairs) + 4(# of G-C pairs)
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DNA and DNA-RNA hybrids are destabilized by....(4)
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1)decr salts concentrations
2)incr detergent concentrations 3)incr temperatures 4)sequence mismatches |
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Then why use a 20 base primer for PCR?
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increases specificity and gives it a higher Tm than if using a smaller primer
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Type of DNA polymerase used in PCR and why?
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Taq DNA Polymerase b/c it is thermally stable
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Steps in PCR (5)
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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 |
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# of copies of amplified DNA in PCR can be determined by this equation....
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(2^N)(y)
N= # of cycles y=starting # of DNA templates |
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Southern Blot
a)use b)requires...(2) b)who invented it |
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 |
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Gel electrophoresis def.
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movement of an electrically charged substance under the influence of an electric field
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Gel electrophoresis steps for DNA (4)
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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. |
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Northern blot difference from Southern blot (4)
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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 |
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Microarrays
a)def. b)what is a microarray c)reverses.... |
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 |
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____ does photolithograph
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GeneChip
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Sample isolation and labeling for Microarray (2)
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1)mRNA is isolated and converted to cDNA via reverse transcriptase
2)cDNA is then fluorescently labeled to allow detection on the array |
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Microarray Applications (5)
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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 |
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Genetic source material for PCR
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DNA
cDNA |
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Genetic source material for DNA Sequencing
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DNA
cDNA |
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Genetic source material for Nucleic Acid Hydridization (north/south blots)
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DNA
RNA cDNA NOT usually used but can be |
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Genetic source material for Microarray (and name of analysis when diff material is used)
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DNA (genomic analysis)
cDNA (transcriptional analysis) |