PCR
What are the components of the Master Mix in the PCR process?
Nucleotides (dATP, dCTP, dGTP, dTTP)
Magnesium ions (enzyme cofactor)
Buffer, containing salt
Taq polymerase
Name the nine signaling pathways
1. RAS-RAF activation of ERK-MRK (MAP Kinase)
2. JAK-STAT
3. Activation of NF-kB
4. HedgeHog
5. Wnt
6. Notch
7. TGF (Transforming Growth Factors)
8. BMP (Bone Morphogenesis Protein)
9. FGF (Fibroblast Growth Factor)
What are three factors that increase mitochondrial damage?
1. Increase ROS
2. Plasma membrane ATP pumps fail
3. Ca ions enter ---> activate enzymes to increase mitochondrial permeability ---> Cytochrome C escapes
What binds to all cell receptors?
LIGANDS
What functions as a ubiquitous intracellular mediator?
Ca2+
How does PCR work?
1.denaturation of the template into single strands, 2.annealing of primers to each original strand for new strand synthesis
3. extension of the new DNA strands from the primers
What determines the speed of response in cell signaling?
The turnover of signaling molecules
Example of responses in cell signaling?
Increase transcription of a gene
alter metabolism
opening an ion channel
Receptor Tyrosine Kinase involve what?
insulin and growth factors
What is Apoptosis?
death of cells
What are the temperatures needed to perform PCR and what happens during each of those temperatures?
Heat (94 C) to denature DNA Strands
Cool (60 C) to anneal primers to template
Warm (72 C) to activate Taq polymerase, which extends primers and replicates DNA
What are the three major classes of Cell-Surface receptor proteins?
Ion-Channel receptor
G-Protein Coupled receptor
E- coupled receptor
How does a mutation in AKT lead to cancer?
stimulating downstream signaling
List the subunits of G-protein?
What mediates most of the effects of Cyclic AMP
(PKA) Cyclic-AMP-dependent Protein Kinase
Where is PV92 Alu located?
Chromosome 16
True or False positive feedback can generate both an all-or-none response?
True
What is the role of nitric oxide as relation to smooth muscle?
Regulates blood flow through by activating soluble guanylate cyclase in the smooth muscle
G Protein-Coupled receptor involve what?
Hormones, neurotransmitters, local mediators, photons, and diverse
What does smell and vision depend on
GCPRs that regulate ion channels
PV92 Alu is dimorphic (meaning two outcomes), what are these outcomes?
641 bp and 941 bp
What enzyme does G-protein attach to?
Adenylate cyclase
Two types of Ligands: what are they and what do they do?
Water-soluble ligands - Glucagon, cannot pass through membrane, recognized by receptor at plasma membrane
Lipid-soluble ligands- can pass through plasma membrane, can diffuse directly across the cell surface membrane. Bind to intracellular receptors in the cytoplasma or nucleus. (Example- Steroids and oestrogen)
Cytokine receptors (JAK) involve what?
immune function, blood cell production, inflammation and tissue repair
What does PI-3-kinase produce and where?
Lipid docking sites in the plasma membrane