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37 Cards in this Set
- Front
- Back
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What is FRAP and what is it used for? |
Fluorescence Recovery After Photobleaching shows protein movement and rate of movement |
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What is FRET and what is it used for? |
Fluorescence Resonance Energy Transfer shows whether two proteins interact in cell |
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What are the purines? |
A, G |
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What are the pyrimidines? |
C, T, U |
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nucleoside vs nucleotide |
nucleoside: sugar and base nucleotide: sugar, base, phosphate, polarity |
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primary structure |
AA sequence, starts with methionine |
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Secondary Structure |
arrangement of AAs in localized regions, stabilized by H bonds Alpha helix and Beta sheets |
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Tertiary Structure |
Folding into functional structure connect alpha helices and beta sheets by loop regions |
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Quaternary Structure |
association of different polypeptide chains |
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Which AAs can by phosphorylated? |
Serine, Threonine, Tyrosine |
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What are amphipathic molecules? |
they have hydrophobic and hydrophilic groups, so molecule turns towards or away from water |
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Which phospholipids are on outside of membrane? |
sphingomyelin and phosphatidylcholine |
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which phospholipids give inside of cell negative charge? |
phosphatidylserine and phosphatidylinositol (b/c have charged head groups) |
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What are phospholipids? |
2 fatty acids and phosphate group linked to glycerol |
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what clusters in lipid rafts? |
sphingomyelin and cholesterol |
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What are GPI anchors made of? |
2 fatty acid side chains, oligosaccharide and ethanolamine |
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How many AAs make up an alpha helix? |
20-25 |
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How many alpha helices do transmembrane proteins have? |
1-5 |
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What are beta barrels? |
transmembrane domains formed by beta sheets highly permeable channels example: Porrins |
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What are the three types of lipid anchors? |
Myristic acid (attach to N terminus) Prenyl Groups Palmitic Acid (last two both added to cysteine residues) |
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what does glycosylation do? |
helps proper folding, stability, and cell-cell adhesion add weight to proteins |
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What does the Nernst Equation describe? |
The relationship between ion concentration and membrane potential, for one ion |
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What is DNA ligase? |
an enzyme that covalently ligates DNA strands |
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What is Antisense RNA/ssDNA used for? |
interfering with gene function --> use synthetic RNA that hybridizes with normal mRNA and blocks translation, then transfection into cultured cells |
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What are the steps of RNAi? |
generate siRNA dsDNA cleaved by dicer aassociates with RISC siRNA unwinds pairs with target mRNA mRNA cleavage translation inhibited |
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Dominant Negative Proteins |
-used to compare wild type and mutant at the same time, -introducing mutant protein directly inhibits protein function |
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Most common method for generating knockout mice? |
In vivo knockout using homologous recombination |
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What is CRISPR/Cas 9 used for and how does it work? |
to inactivate/mutate a gene Cas 9 cuts DNA and recognizes guide RNAs which take it to the DNA |
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What are DNA microarrays used for? |
to find out what's changed in a cell type |
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what is southern blotting used for? |
DNA- to confirm whether homologous recombination has occurred |
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what is RNA-seq for, and what is it? |
allows you to find out frequency of mRNAs/nucleotides ---cellular mRNAs reverse transcribed to cDNAs |
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what does in situ hybridization tell you? |
where and how much RNA transcript is being expressed |
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What does Western Blotting do? |
allows you to visualize protein from a cell extract by detecting bound antibody semi-quantitative |
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What is x-ray crystallography used for? |
to show size and structure of DNA provided evidence for semiconservative replication (14N and 15N media for DNA) |
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What is immunocytochemistry used for? |
To visualize where proteins are in an intact cell |
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What is Immunoprecipitation used for? |
to detect the physical interactions between two proteins -use insoluble beads to collect complexes |
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What is the Yeast Two-Hybrid System used for? |
to figure out what interacts with protein of interest (interactions between pairs of proteins in yeast cells) |
