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106 Cards in this Set
- Front
- Back
Mitochondria |
Cellular respiration to produce ATP
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Endosymbiont Theory |
primitive bacteria formed symbiotic relationship with early eukaryotic cells
gradual transfer of mt genes to nucleus |
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Mitochondrial Inheritance |
Extranuclear = non Mendelian = maternal = uniparental
offspring have phenotype of mother in mt genes
sperm does not contribute bc mt is lost after fertilization |
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Neurospora |
Mt Mutation:
[Poky] mutation - strains grow slowly for days, then rate accelerates |
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Leber's Hereditary Optic Neuropathy (LHON) |
adult optic nerve degeneration
electron transport chain mutation |
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Heteroplasmy |
mixed population of mt in a cell
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Mt Genome |
Circular (usually) -37 genes, have own ribosomes
Some proteins required for these are encoded by nuclear genes |
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E. coli |
Lower intestines of animals
pathogenic strains
have singular circular chromosome |
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Auxotroph |
nutritional mutant - requires one or more supplements to grow EX: AA's |
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Symbols |
Amp^r is resistance Str^s is sensitivity Auxotroph needs this - |
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E. coli Biology |
Prokaryote - nucleoid region contains the chromosome
Binary Fisson -> Exponential Growth |
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Colony |
visible cluster of clones of bacteria
- about 1 million cells per unit
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Lawn |
entire plate covered, no individual colonies |
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Lag Phase |
slow or no apparent bacterial growth |
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Log Phase |
bacteria double every 20 min to 1 x 10^9/ml |
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Stationary Phase |
nutrient and/or oxygen limited cell number remember constant |
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Minimal Media |
essentials for organisms - Sugar (carbon source) + salts - bacteria synth AA, nucleotides and vitamins |
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Selective Media |
Allows one species to grow while selecting against another |
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Phototrophs |
organism synths requirements from minimal media |
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Conditional Mutant |
mutation is expressed in a certain condition
temp sensitive mutant can function at 25 C but not at 42 C |
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Resistance Mutant |
antibiotic resistance in bacteria
Amp^r |
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Conjugation |
aka parasexual mating - one way transfer of genetic info |
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Conjugation |
aka parasexual mating - one way transfer of genetic info |
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Plasmid |
circular, episomally maintained DNA |
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Conjugation |
aka parasexual mating - one way transfer of genetic info |
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Plasmid |
circular, episomally maintained DNA |
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Conjugation |
1. Pilus attaches to cell 2. nick DNA -> transfer DNA 3. DNA pol makes dsDNA 4. break pilus |
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Knockout Mouse Creation |
1. Prepare construct DNA in lab 2. Add it to embryo in culture 3. Homologous Rec by cell 4. Implant in surrogate 5. Cross breed to make knockout |
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Transformation |
naked DNA enters cell, brings new genes
bacteria with new DNA is a transformant |
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Competent Cells |
cells made to be able to take on DNA - CaCl2 treat bacteria to make cell membrane permeable to naked DNA |
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Competent Cells |
cells made to be able to take on DNA - CaCl2 treat bacteria to make cell membrane permeable to naked DNA |
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Plasmid Requirements |
1. Must have an Ori (for replication) 2. Selectable Marker 3. Restriction Enzyme Sites 4. High copy number in E. coli |
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Transduction |
Bacteriophage is virus that infects bacteria 1. Adsorbs to cell and injects DNA 2. Normal activity stops, becomes "phage factory" 3. Host DNA broken to pieces, new viruses released to other cells
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Competent Cells |
cells made to be able to take on DNA - CaCl2 treat bacteria to make cell membrane permeable to naked DNA |
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Plasmid Requirements |
1. Must have an Ori (for replication) 2. Selectable Marker 3. Restriction Enzyme Sites 4. High copy number in E. coli |
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Transduction |
Bacteriophage is virus that infects bacteria 1. Adsorbs to cell and injects DNA 2. Normal activity stops, becomes "phage factory" 3. Host DNA broken to pieces, new viruses released to other cells
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Faulty Head Stuffing |
piece of bacteria's DNA packaged into a virus
- the phage infects new cell and transfers genes from the first bacterium |
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Lysogenic Pathway |
Phage DNA enters specific location in chromosome
prophage integrates and is lysogenic
phage gene represses lytic cycle |
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Virulent Phage |
phage that is always lytic |
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Virulent Phage |
phage that is always lytic |
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Temperate Phage |
phage is always lysogenic |
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Viral Gene Therapy |
goal is to insert the normal gene into human DNA while using a virus as the vector |
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VGT Example |
Treat a severe immune deficiency (ADA) 1. Remove viral replication genes and insert normal ADA gene into virus 2. Remove T cells from patient 3. Infect cells with engineered virus 4. Infuse into patient |
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Problems with Gene Therapy |
inflammatory response to the engineered virus can lead to death gene could disrupt cell cycle gene leading to cancer |
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Bacterial Genes |
Many are constitutively expressed - "housekeeping" genes Others are regulated - can be turned on/off depending on cell needs |
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Operon |
group of coordinately regulated genes - 1 promoter for a number of genes - Polycistronic mRNA (containing many cistrons, or genes) |
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Lac Operon |
in the presence of lactose: -cells convert lactose to glucose and galactose - lactose is an inducer of this (also an effector) - inducible |
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Lac Z Gene |
encodes betagalactosidase b-gal lactose ------> glucose + galactose bgal flourishes in presence of lactose |
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Lac Y Gene |
encodes permease that transports lactose into cells |
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Lac Operon |
normal repressed because lactose is absent the operon is induced when lactose is present |
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Lac 1 Gene |
encodes a repressor protein
- repressor binds to operator |
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hGH deficiency |
Mutation in hGH gene --> drawfism |
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Making cDNA Library |
1. Start with mRNA - cDNA is tissue specific - cDNA has no introns 2. Reverse transcriptase enzyme copies mRNA to cDNA 3. Restriction enzyme digest the cDNAs to obtain "sticky ends" 4. Ligate cDNAs into plasmid vectors 5. Transform bacteria with the plasmids 6. Pick bacterial colony with hGH plasmid 7. Grow it and it will express the hGH gene |
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Transgenic Plants |
Plants acquire new genetic trait by direct introduction of foreign gene 1. Agrobacterium carries plasmid and infects plant cell 2. Gene gun shoots DNA into plant cells |
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Benefits of Transgenic Plants |
disease/insect resistance herbicide/drought tolerance repress allergens increase nutrition |
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Drawbacks of Transgenic Plants |
increased seed costs pesticide resistant bugs harmful to insects unknown |
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Bt Corn |
Bacillus thuringiensis bacteria produce toxic protein - Bt protein binds to receptors in insect intestine and blocks feeding |
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Transgenic Goats |
produce human protein (drug) in milk ex: Human EPO - mammary gland-specific promoter - beta lactoglobulin promoter |
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Microinjection |
1. Inject gene construct into fertilized egg, integrates into chromosome 2. Implant embryo into surrogate mother 3. Transgenic goat produces human drug 4. Purify protein from milk |
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Examples of Transgenic Animals |
Agriculture- pigs made to digest more and produce less manure Medicine- chicken produces an antibody in eggs Fish Farming- engineered salmon grow faster |
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Xenotranslplantation |
pigs have smaller sized organs to humans - knockout pig cell surface antigens to prevent hyperacute rejections |
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Patenting |
Raw products of nature are not patentable - you can patent a gene, method, or animal |
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Gene, Reproductive and Therapeutic |
3 types of cloning |
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Gene Cloning |
recombiant bacteria - transgenic plants and animals |
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Reproductive Cloning |
Yields an organism - embryo twinning or nuclear transfer |
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Therapeutic Cloning |
nuclear transfer for stem cells to treat disease - isolate nucleus from a somatic cell - fuse with enucleate donor egg |
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Embryo Twinning |
1 sperm + 1 egg = 2 embryos (genetically identical) |
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Somatic Cell Nuclear Transfer |
obtain somatic cell from donor cell must be in Go stage of interphase fuse somatic cell nucleus with enucleate egg - grow embryo for 6 days, implant into surrogate mother |
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Ethical Implications of SCNT |
human cloning "playing with nature?" social challenges for the clones regulation |
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Totipotent Stem Cells |
can differentiate into any cell type including placenta ex: early embryo |
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Pluripotent Stem Cells |
ICM can differentiate into any body cell type ex: 5 day old embryo |
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Multipotent Stem Cells |
give rise to a number of cell types ex: cells in bone marrow |
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Blastocyst |
structure formed in early development of mammals - contains inner cell mass (future embryo) |
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Inner Cell Mass |
the mass of visible cells in the blastocyst that gives rise to an embryo |
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Germ-Line Mutation |
In sperm or egg, mutation will be passed to offspring |
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Somatic Mutation |
body cell mutations, not passed to offspring |
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Spontaneous Mutation |
mutation is a natural aspect of DNA replication |
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Induced Mutation |
mutation caused by UV light, chemicals, x-ray, etc. |
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Point Mutations |
mutations that involve single base substitution, deletion addition |
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Transition (Base Pair Substitution) |
G to A (purines) or T to C (pyrimidines) |
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Transversion (Base Pair Substitution) |
G to C, G to T OR A to T, A to C (vice versa) |
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Nonsense Mutation |
premature stop codon - Codon AAA --> UAA lys stop |
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Silent Mutation |
mutation with generally no amino acid change - no effect on organism |
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Frameshift Mutation |
mutation that shifts the reading frame - may lead to a premature stop codon |
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Fragile X Syndrome |
CGG repeats more than 200 times, results in a weakened X chromosome - causes FMR1 gene splicing by hypermethylation |
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Small Deletion: Cystic Fibrosis |
CFTR gene encodes a chloride channel active in cells that line the lungs |
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Xeroderma pigmentosum |
Autosomal recessive, extreme sun sensitivity
- high risk of skin cancer, corneal damage |
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Cancer |
abnormal cells divide uncontrollably and destroys body tissue |
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Tumor |
neoplasm, mass of cells |
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Benign |
cells in a single, contained mass |
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Malignant |
invades the surrounding tissue
- cells may break off and move (metastasis) |
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Transformed Cell |
a cell that has lost normal growth controls - loses contact inhibition - immortal |
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Proto-oncogenes |
the normal genes involved in cell cycle - encode proteins (growth factors and receptors) - if it mutates, becomes oncogene, leads to uncontrolled growth |
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Oncogenesis |
when a protooncogene mutates to an oncogene - initiation of cancer |
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Ras |
signal transducer for cell division |
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Tumor Suppressor Genes |
protein products suppress uncontrolled cell proliferation
- both copies must be inactivated for loss of function (2 mutations in one cell required) |
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pRB tumor suppressor protein |
allows cell to progress to S phase - a G1 to S checkpoint protein |
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p53 tumor supressor |
involved in ~50% of cancers - monitors signals that indicate DNA damage - damage cell --> increase p53 |
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Steps of p53 repair |
1. Damage DNA 2. p53 3. DNA repair, cell cycle arrest or apoptosis 4. Genome integrity |
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Apoptosis |
programmed cell death - p53 can activate this pathway |
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Cyclins |
proteins that control the cell cycle |
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Cdks |
cyclin dependent kinases
- complex with cyclins - phosphorylate other molecules - G1 to S checkpoint |
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Cancer is a Multi-Step Disease |
accumulation of mutations in a number of genes in a single cell - can build up over decades |
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Genetic Predisposition |
inherit one or more mutant alleles, increases the risk of cancer |
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Retinoblastoma |
eye cancer develops in childhood
- hereditary or sporadic - child born RB/RB in all cells
Sporadic- develop 2 mutations in 1 cell after birth - only find mutation in eye cells, not blood |
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Types of Cancer |
Carcinoma, 90% of cancers Sarcomas, rare Lukemia and Lymphomas, 8% of tumors |