Genetics Final

Front 1

Mitochondria

Back 1

Cellular respiration to produce ATP

Front 2

Endosymbiont Theory

Back 2

primitive bacteria formed symbiotic relationship with early eukaryotic cells

gradual transfer of mt genes to nucleus

Front 3

Mitochondrial Inheritance

Back 3

Extranuclear = non Mendelian = maternal = uniparental

offspring have phenotype of mother in mt genes

sperm does not contribute bc mt is lost after fertilization

Front 4

Neurospora

Back 4

Mt Mutation:

[Poky] mutation

- strains grow slowly for days, then rate accelerates

Front 5

Leber's Hereditary Optic Neuropathy (LHON)

Back 5

adult optic nerve degeneration

electron transport chain mutation

Front 6

Heteroplasmy

Back 6

mixed population of mt in a cell

Front 7

Mt Genome

Back 7

Circular (usually)

-37 genes, have own ribosomes

Some proteins required for these are encoded by nuclear genes

Front 8

E. coli

Back 8

Lower intestines of animals

pathogenic strains

have singular circular chromosome

Front 9

Auxotroph

Back 9

nutritional mutant

- requires one or more supplements to grow

EX: AA's

Front 10

Symbols

Back 10

Amp^r is resistance

Str^s is sensitivity

Auxotroph needs this -

Front 11

E. coli Biology

Back 11

Prokaryote

- nucleoid region contains the chromosome

Binary Fisson -> Exponential Growth

Front 12

Colony

Back 12

visible cluster of clones of bacteria

- about 1 million cells per unit

Front 13

Lawn

Back 13

entire plate covered, no individual colonies

Front 14

Lag Phase

Back 14

slow or no apparent bacterial growth

Front 15

Log Phase

Back 15

bacteria double every 20 min to 1 x 10^9/ml

Front 16

Stationary Phase

Back 16

nutrient and/or oxygen limited

cell number remember constant

Front 17

Minimal Media

Back 17

essentials for organisms

- Sugar (carbon source) + salts

- bacteria synth AA, nucleotides and vitamins

Front 18

Selective Media

Back 18

Allows one species to grow while selecting against another

Front 19

Phototrophs

Back 19

organism synths requirements from minimal media

Front 20

Conditional Mutant

Back 20

mutation is expressed in a certain condition

temp sensitive mutant can function at 25 C but not at 42 C

Front 21

Resistance Mutant

Back 21

antibiotic resistance in bacteria

Amp^r

Front 22

Conjugation

Back 22

aka parasexual mating

- one way transfer of genetic info

Front 23

Conjugation

Back 23

aka parasexual mating

- one way transfer of genetic info

Front 24

Plasmid

Back 24

circular, episomally maintained DNA

Front 25

Conjugation

Back 25

aka parasexual mating

- one way transfer of genetic info

Front 26

Plasmid

Back 26

circular, episomally maintained DNA

Front 27

Conjugation

Back 27

1. Pilus attaches to cell

2. nick DNA -> transfer DNA

3. DNA pol makes dsDNA

4. break pilus

Front 28

Knockout Mouse Creation

Back 28

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

Front 29

Transformation

Back 29

naked DNA enters cell, brings new genes

bacteria with new DNA is a transformant

Front 30

Competent Cells

Back 30

cells made to be able to take on DNA

- CaCl2 treat bacteria to make cell membrane permeable to naked DNA

Front 31

Competent Cells

Back 31

cells made to be able to take on DNA

- CaCl2 treat bacteria to make cell membrane permeable to naked DNA

Front 32

Plasmid Requirements

Back 32

1. Must have an Ori (for replication)

2. Selectable Marker

3. Restriction Enzyme Sites

4. High copy number in E. coli

Front 33

Transduction

Back 33

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

Front 34

Competent Cells

Back 34

cells made to be able to take on DNA

- CaCl2 treat bacteria to make cell membrane permeable to naked DNA

Front 35

Plasmid Requirements

Back 35

1. Must have an Ori (for replication)

2. Selectable Marker

3. Restriction Enzyme Sites

4. High copy number in E. coli

Front 36

Transduction

Back 36

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

Front 37

Faulty Head Stuffing

Back 37

piece of bacteria's DNA packaged into a virus

- the phage infects new cell and transfers genes from the first bacterium

Front 38

Lysogenic Pathway

Back 38

Phage DNA enters specific location in chromosome

prophage integrates and is lysogenic

phage gene represses lytic cycle

Front 39

Virulent Phage

Back 39

phage that is always lytic

Front 40

Virulent Phage

Back 40

phage that is always lytic

Front 41

Temperate Phage

Back 41

phage is always lysogenic

Front 42

Viral Gene Therapy

Back 42

goal is to insert the normal gene into human DNA while using a virus as the vector

Front 43

VGT Example

Back 43

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

Front 44

Problems with Gene Therapy

Back 44

inflammatory response to the engineered virus can lead to death

gene could disrupt cell cycle gene leading to cancer

Front 45

Bacterial Genes

Back 45

Many are constitutively expressed

- "housekeeping" genes

Others are regulated

- can be turned on/off depending on cell needs

Front 46

Operon

Back 46

group of coordinately regulated genes

- 1 promoter for a number of genes

- Polycistronic mRNA (containing many cistrons, or genes)

Front 47

Lac Operon

Back 47

in the presence of lactose:

-cells convert lactose to glucose and galactose

- lactose is an inducer of this (also an effector)

- inducible

Front 48

Lac Z Gene

Back 48

encodes betagalactosidase

b-gal

lactose ------> glucose + galactose

bgal flourishes in presence of lactose

Front 49

Lac Y Gene

Back 49

encodes permease that transports lactose into cells

Front 50

Lac Operon

Back 50

normal repressed because lactose is absent

the operon is induced when lactose is present

Front 51

Lac 1 Gene

Back 51

encodes a repressor protein

- repressor binds to operator

Front 52

hGH deficiency

Back 52

Mutation in hGH gene --> drawfism

Front 53

Making cDNA Library

Back 53

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

Front 54

Transgenic Plants

Back 54

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

Front 55

Benefits of Transgenic Plants

Back 55

disease/insect resistance

herbicide/drought tolerance

repress allergens

increase nutrition

Front 56

Drawbacks of Transgenic Plants

Back 56

increased seed costs

pesticide resistant bugs

harmful to insects

unknown

Front 57

Bt Corn

Back 57

Bacillus thuringiensis bacteria produce toxic protein

- Bt protein binds to receptors in insect intestine and blocks feeding

Front 58

Transgenic Goats

Back 58

produce human protein (drug) in milk

ex: Human EPO

- mammary gland-specific promoter

- beta lactoglobulin promoter

Front 59

Microinjection

Back 59

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

Front 60

Examples of Transgenic Animals

Back 60

Agriculture- pigs made to digest more and produce less manure

Medicine- chicken produces an antibody in eggs

Fish Farming- engineered salmon grow faster

Front 61

Xenotranslplantation

Back 61

pigs have smaller sized organs to humans

- knockout pig cell surface antigens to prevent hyperacute rejections

Front 62

Patenting

Back 62

Raw products of nature are not patentable

- you can patent a gene, method, or animal

Front 63

Gene, Reproductive and Therapeutic

Back 63

3 types of cloning

Front 64

Gene Cloning

Back 64

recombiant bacteria

- transgenic plants and animals

Front 65

Reproductive Cloning

Back 65

Yields an organism

- embryo twinning or nuclear transfer

Front 66

Therapeutic Cloning

Back 66

nuclear transfer for stem cells to treat disease

- isolate nucleus from a somatic cell

- fuse with enucleate donor egg

Front 67

Embryo Twinning

Back 67

1 sperm + 1 egg = 2 embryos (genetically identical)

Front 68

Somatic Cell Nuclear Transfer

Back 68

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

Front 69

Ethical Implications of SCNT

Back 69

human cloning "playing with nature?"

social challenges for the clones

regulation

Front 70

Totipotent Stem Cells

Back 70

can differentiate into any cell type including placenta

ex: early embryo

Front 71

Pluripotent Stem Cells

Back 71

ICM can differentiate into any body cell type

ex: 5 day old embryo

Front 72

Multipotent Stem Cells

Back 72

give rise to a number of cell types

ex: cells in bone marrow

Front 73

Blastocyst

Back 73

structure formed in early development of mammals

- contains inner cell mass (future embryo)

Front 74

Inner Cell Mass

Back 74

the mass of visible cells in the blastocyst that gives rise to an embryo

Front 75

Germ-Line Mutation

Back 75

In sperm or egg, mutation will be passed to offspring

Front 76

Somatic Mutation

Back 76

body cell mutations, not passed to offspring

Front 77

Spontaneous Mutation

Back 77

mutation is a natural aspect of DNA replication

Front 78

Induced Mutation

Back 78

mutation caused by UV light, chemicals, x-ray, etc.

Front 79

Point Mutations

Back 79

mutations that involve single base substitution, deletion addition

Front 80

Transition (Base Pair Substitution)

Back 80

G to A (purines) or T to C (pyrimidines)

Front 81

Transversion (Base Pair Substitution)

Back 81

G to C, G to T OR A to T, A to C (vice versa)

Front 82

Nonsense Mutation

Back 82

premature stop codon

- Codon AAA --> UAA

lys stop

Front 83

Silent Mutation

Back 83

mutation with generally no amino acid change

- no effect on organism

Front 84

Frameshift Mutation

Back 84

mutation that shifts the reading frame

- may lead to a premature stop codon

Front 85

Fragile X Syndrome

Back 85

CGG repeats more than 200 times, results in a weakened X chromosome

- causes FMR1 gene splicing by hypermethylation

Front 86

Small Deletion: Cystic Fibrosis

Back 86

CFTR gene encodes a chloride channel active in cells that line the lungs

Front 87

Xeroderma pigmentosum

Back 87

Autosomal recessive, extreme sun sensitivity

- high risk of skin cancer, corneal damage

Front 88

Cancer

Back 88

abnormal cells divide uncontrollably and destroys body tissue

Front 89

Tumor

Back 89

neoplasm, mass of cells

Front 90

Benign

Back 90

cells in a single, contained mass

Front 91

Malignant

Back 91

invades the surrounding tissue

- cells may break off and move (metastasis)

Front 92

Transformed Cell

Back 92

a cell that has lost normal growth controls

- loses contact inhibition

- immortal

Front 93

Proto-oncogenes

Back 93

the normal genes involved in cell cycle

- encode proteins (growth factors and receptors)

- if it mutates, becomes oncogene, leads to uncontrolled growth

Front 94

Oncogenesis

Back 94

when a protooncogene mutates to an oncogene

- initiation of cancer

Front 95

Ras

Back 95

signal transducer for cell division

Front 96

Tumor Suppressor Genes

Back 96

protein products suppress uncontrolled cell proliferation

- both copies must be inactivated for loss of function (2 mutations in one cell required)

Front 97

pRB tumor suppressor protein

Back 97

allows cell to progress to S phase

- a G1 to S checkpoint protein

Front 98

p53 tumor supressor

Back 98

involved in ~50% of cancers

- monitors signals that indicate DNA damage

- damage cell --> increase p53

Front 99

Steps of p53 repair

Back 99

1. Damage DNA

2. p53

3. DNA repair, cell cycle arrest or apoptosis

4. Genome integrity

Front 100

Apoptosis

Back 100

programmed cell death

- p53 can activate this pathway

Front 101

Cyclins

Back 101

proteins that control the cell cycle

Front 102

Cdks

Back 102

cyclin dependent kinases

- complex with cyclins

- phosphorylate other molecules

- G1 to S checkpoint

Front 103

Cancer is a Multi-Step Disease

Back 103

accumulation of mutations in a number of genes in a single cell

- can build up over decades

Front 104

Genetic Predisposition

Back 104

inherit one or more mutant alleles, increases the risk of cancer

Front 105

Retinoblastoma

Back 105

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

Front 106

Types of Cancer

Back 106

Carcinoma, 90% of cancers

Sarcomas, rare

Lukemia and Lymphomas, 8% of tumors