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58 Cards in this Set
- Front
- Back
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Basic structure of a nucleotide
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Nitrogenous base
Ribose Phosphate |
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Difference between DNA & RNA
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DNA
--Deoxiribose --AGCT --Double helix RNA --Ribose --AGCU --Single stranded or w/ limited folding |
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Biology's Central Dogma
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DNA -xscrpn--> RNA -xlatn--> protein
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Most common gene in the human genome
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Alu-I
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What are the specifics of Alu-I
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-Most common gene in human genome
-7% of DNA in cells is AluI -It is a 'Retrotransposon' -Makes two proteins ----Reverse transcriptase ----Transposases (cut/paste) |
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Transposon
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Known as a "jumping gene"
sequences of DNA that can move or transpose themselves to new positions within the genome of a single cell. |
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Retrotransposon
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Genes that move by a "copy and paste" mechanism, the copy is made of RNA, not DNA.
Genetic elements that can amplify themselves in a genome and are ubiquitous components of the DNA of many eukaryotic organisms. |
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Reverse Transcription
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RNA --> DNA
The process of creating double-stranded DNA from a single-stranded RNA template (the reverse of the standard process) |
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What is the secondary structure of DNA?
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An anti-parallel double-helix
((3' end of one strand faces the 5' end of its counterpart)) |
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Griffith's experiment
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1928 - of the first experiments suggesting that bacteria are capable of transferring genetic information through a process known as transformation.
Infect mice with Smooth/Rough bacteria: Rough - mouse lives Smooth - mouse dies Heat killed smooth - mouse lives Rough and H.K. smooth - mouse dies. Live S found in blood of dead mice ((there is some 'transforming principle')) |
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Avery, MacLeod, McCarty's experiment
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1944 - experiment demonstration that DNA is the substance that causes bacterial transformation.
-Strains of pneumococcus smooth (S) bacteria in test tubes (not mice) -Detergent to burst heat-killed S cells -Separates in tubes into cell debris and lysate (cell coat, protein, dna, rna) -Use 3 samples --1. S-Lysate (S-guts: cell coat, protein, dna, rna) --2. R strain --3. Lysate + R strain Test 1 Mix tube of S-lysate w/ enzyme to remove S-cell sugar walls from lysate; mix with R-cells - Cells transformed to S-cells; Sugar wall not transforming principle. Test 2 Add protein digesting enzymes to S-lysate; mix with R strain; Still transformed to S so proteins not transforming principle! Test 3 Precipitate from S-lysate, the nucleic acids, DNA/RNA, with alcohol; dissolved precipitate in water.... Destroy RNA w/ RNase enzyme - the R-strain transformed to S; its not RNA either! Test 4 Used DNase enzyme to destroy DNA; mix with R-cells; the solution did not transform - they remained R-cells. DNA is the transform |
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Hershey/Chase experiment
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1952 -confirmation that DNA is the genetic material that carried the information for inheritance.
Used radio-labeled molecules (P32 and S35) to trace. S35 added to phage, bind to cell wall of cell, injects DNA - blend/centrifuge - S35 found in bottom w/ heavier, non-DNA material. P32 added to phage, bind to cell wall of cell, injects DNA - blend/centrifuge - P32 found in area of medium weight DNA material. |
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Introns - purpose for
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- Alternate splicing
- Protection against retrotransposon events |
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Gene
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Unit of heredity
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Allele
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A gene variant
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Genotype
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Allelic makeup of an individual
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Genome
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Complete genetic makeup of an organism
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Phenotype
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Physical characteristic
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Homozygous
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A gene with identical alleles for that gene
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PCR
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PCR is polymerase chain reaction
Used to amplify (multiply) small amounts of DNA in order to work with a large sample of DNA. |
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Mechanisms of PCR
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Denature-->Anneal-->Elongation
Denature -Heats to melt DNA Anneal -Slowly cool so primers bond to single-stranded DNA (DNA synthesis begins) Elongate -Raise temp to optimal so DNA (Taq) Polymerase can synthesize a complimentary strand of DNA |
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How can we determine if an organism has a particular gene?
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PCR (Polymerase chain reaction)
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Electrophoresis
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Used for separating DNA molecules of different sizes.
Small fragments of duplex DNA in slots in one end of gel. Submerged in buffer and subjected to electric field. DNA moves in response to electric field with accordance to their lengths. Black and Blue go to Red |
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Process to genetically engineer a fish
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1a. Extract DNA from Chinook
1b. PCR to amplify growth gene 2a. Extract DNA from Prout 2b. PCR to amplify promoter 3. Ligation Reaction (promoter+gene) 4. PCR to amplify fragments Insert gene into the Atlantic Salmon - Use a virus - Gene gun - Micro-injection |
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How are genes inserted into a foreign organism?
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1. Use a virus
2. Gene gun 3. Micro-injection |
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Draw a nucleotide (dNTP)
where N=A, T, C, or G |
Cyclops, missing left foot, holding base in left hand and P in right hand (torch)
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Draw dAMP
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Remove two phosphates from dNTP
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Ingredients needed to do PCR
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*Template DNA
*Primers (Two, DNA fragments, directional) *Taq DNA polymerase *Nucleotides (dNTP) Buffer MgCl2 Water |
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Molecules for Life
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*Nucleotides (RNA/DNA)
-Protein -Lipids -Carbohydrates |
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Given any base, what might cause replication to stop?
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Missing a 3' hydroxyl group
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Who developed PCR
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Kary Mullis (surfer LSD Nobel guy)
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Procedure for extracting DNA
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-Break intercellular bonds
-Soap to break down lipid bilayer/nucleus walls -Filter gunk out of solution -Alcohol to precipitate DNA -Salt neutralizes DNA charge, removes proteins -Ice deactivates enzymes that break down DNA, AND speeds precipitation of nucleic acids |
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Three methods of bacterial sex
(and who doesn't like watching bacterial sex??) |
(Transformation)
Taking genes from the environment and increasing diversity (Conjugation) Transferring plasmid from f+ to f- (Transduction) Process by which DNA is transferred from one bacterium to another via a virus. Bacteriophage injects DNA into a host cell - two things may occur: 1) Lytic Cycle -Expressed genes burst out of cell and infect 2) Lysogenic phage chromosome is integrated into the bacterial chromosome where it remains dormant; genome is excised when under stress - then goes into Lytic Cycle. |
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Incomplete Dominance
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Blending
Phenotype of heterozygous genotype is intermediate between the phenotypes of the homozygous genotypes. |
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Co-Dominance
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Alternative forms of a gene can both be detected when they are present in a cell or organism (eg. Blood Type)
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Alternative Splicing
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Exons of the RNA are reconnected in multiple ways
Introns are spliced via two means: 1) Self Splicing 2) Protein Mediated Splicing (splicesome) |
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What happens to genes without introns?
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If large enough, they are detected in the nucleous and disabled.
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Methylation
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Where RIP (repeat induced point mutations) occur to disable genes -
Cytosine (pyramidine) converted to Thiamines - disabling the gene. |
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Pseudo Gene
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A methylated gene
Also known as a 'fossil gene' - contain a large number of T (thiamine) due to methylation. |
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Repeat Induced Point-Mutations (RIP)
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Process by which DNA accumulates G:C to A:T mutations. It is associated with DNA methylation. ((Sequences that are longer than a few hundred base pairs and that are repeated in the haploid genome are subjected to RIP.))
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Epistasis
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Any type of interaction in which the genotype at one locus effects the phenotypic expression of the genotype at another locus.
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Penetrance
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#Pheno/Total
The proportion of organisms whose phenotype matches the genotype for a given trait. |
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Incomplete Penetrance
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Genotype is not expressed to any detectable degree in some individuals (The extreme of variable expressivity)
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Principle of Segregation
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Mendell - an individual inherits an allele for a trait from each parent. The alleles segregate from each other when forming gametes.
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Expressivity
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Variations of a phenotype in individuals carrying a particular genotype.
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Independent Assortment
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Mendel - alleles of different genes assort independently of one another during gamete formation.
This means that traits are transmitted to offspring independently of one another. |
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Why is PCR one of the most important molecular techniques?
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DNA studies would be nearly impossible without it.
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PCR: What are primers?
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Short synthetic pieces of DNA for the DNA polymerase to build on.
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Draw the structure of an amino acid
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Amine group
Carboxylic group Amino Acid side chain Amine--CR--Carboxyl |
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Why is a primer needed for sequencing?
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To provide the 3' hydroxyl to allow attachment of the deoxynucleotide for DNA elongation.
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Espressivity
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Variations of a phenotype in individuals carrying a particular genotype
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Why might an organism show P(A_)=2/3 and P(aa)=1/3?
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Because being homozygous for the trait is lethal to the organism (eg. manx cat, never see MM)
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Transformation
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Bacterial reproduction that takes genes from the environment to increase diversity
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Transduction
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Bacterial reproduction
Process by which DNA is transferred from one bacterium to another via a virus. Bacteriophage injects DNA into a host cell - two things may occur: 1) Lytic Cycle -Expressed genes burst out of cell and infect 2) Lysogenic phage chromosome is integrated into the bacterial chromosome where it remains dormant; genome is excised when under stress - then goes into Lytic Cycle. |
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Conjugation
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Bacterial reproduction
Transferring a plasmid from f+ to f- |
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Plasmid
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Extrachromosomal circular DNA
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Homologous
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Gene derived from a common ancestor
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Ploidy
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Number of copies of chromosomes
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