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How do you do a genetic screen for haploid organisms?
temperature sensitive mutants (conditionally defective protein products). displays mutant phenotype w/ temp shift.
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Explain a complementation test
homozygous individual for one recessive mutation mated with one homozygous for other mutation. If offspring are on same gene, offspring will show mutant phenotype.
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Mutagenisis method relies on the fact that exogenous DNA inserted randomly into the genome can produce mutations if the inserted fragment interrupts a gene or its regulatory sequences
insertional mutagenesis
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Segment of DNA that can move from one position in a genome to another
transposon
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linkage analysis
Co-inheritance of two genetic loci that lie near each other on the same chromosome. The closer together the two loci, that is, the greater the linkage, the lower the frequency of recombination between them. The closer the genes are, the greater the likelihood they will be passed to offspring as a pair.
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Technique for analyzing the simultaneous expression of large numbers of genes in cells, in which isolated cellular RNA is hybridized to a large array of short DNA probes immobilized on glass slides. he exact sequence—and position—of every probe on the chip is known. Thus any nucleotide fragment that hybridizes to a probe on the array can be identified as the product of a specific gene simply by detecting the position to which it is bound.
DNA microarrays
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cDNA
DNA molecule made as a copy of messenger RNA and therefore lacking the introns that are present in genomic DNA.
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start with a particular gene and proceed to make mutations in it, creating mutant cells or organisms so as to analyze the gene's function.
reverse genetics
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complementary to vector
sequence flanking insert region (can
be used to sequence all inserts)
“Universal”primers
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Introduction of a foreign DNA molecule into a eucaryotic cell. It is usually followed by expression of one or more genes in the newly introduced DNA.generally leads to gene addition rather than gene replacement: the altered gene inserts at a random location in the genome, so that the cell (or the organism) ends up with the mutated gene in addition to its normal gene copies
transfection
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antisense RNA
RNA complementary to a specific RNA transcript of a gene that can hybridize to the specific RNA and block its function
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Photosynthesis converts ____ and ____ to ____ and ____ with energy from_____
C02 and H2O....O2 and sugars....sunlight
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During respiration, in addition to ouputting H2O and CO2, what else is created?
Chemical bond energy
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Catabolism
and energetically favorable reaction that oxidizes food molecules and exports energy on activated carrier molecules
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Anabolism
An energetically unfavorable reaction in which a molecule available in the cell is converted into a molecule needed by the cell
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null mutation
a loss of function mutation that completely abolishes the activity of the gene
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dominant-negative mutations
dominant acting mutation that blocks gene activity, causing a loss of function phenotype even in the presence of a normal copy of the gene. mutant interferes with the function of the normal gene product.
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suppressor mutation
suppresser the phenotypic effect of another mutation, so that the double mutant seems normal.
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intragenic suppressor mutation
lies within the gene affected by the first mutation
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extragenic suppressor mutation
lies on a second gene from the first mutation, oftenone whose product interacts directly with the products of the first.
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sonicate
to shear DNA into random sized fragments
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adaptor
sequencing end primer
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microarrays use what type of DNA?
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in a microarray, each dot is an/a...
oligo
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most organisms use ____ for gene expression
mRNA
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What can RNAi be used for?
make knock-down genes.
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Overview of Glycolysis
Invest 2 ATPs in coupled reactions, cleave 6 carbon sugar to three two carbon sugars, harvest 4 ATP's and 2 NADH's in coupled reactions. Produce 2 Pyruvate molecules
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Fermentation
degrade 2 pyruvate molecules to carbon dioxide and ethanol.
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name pyruvate's 2 major pathways of anaerobric energy production
- fermentation to ethanol
- frementation to lactic acid
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name pyruvate's 1 major pathway of aerobric energy production
citric acid cycle through acetyl CoA
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pyruvates and fatty acids are converted to acetyl CoA in the...
mitochondrion
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describe the pyruvate dehydrogenase complex
(Pyruvate is converted to Acetyl CoA+ NADH+CO2)X2
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What are two foodmolecule from the cytosol that contribute to reactions in the mitochondrion?
fatty acids and pyruvate
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microtubles eminate from....
centrosomes
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polymer of actin monomers, dimer of alpha and beta
microfilaments
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where does glycolysis occur?
cytosol
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where does the citric acid cycle occur?
mitochondrial matrix
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Where doe oxidative phosphorolation occur?
mitochondrial membrane
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what are the two mitochondrial compartments?
matrix and intermembrane space
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Th matrix of the mitochondri contains:
mixture of hundreds of enzymes including those required for the oxidation of pyruvate and fatty acids for the citric acid cycle
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Describe the inner membrane of the mitochondria
folded into cristae, contains proteins that carry out oxidation reactions of the electron transport chain and the ATP synthase that makes ATP in the matrix
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Describe the outer membrane of mitochondria
contains porin (large, channel-forming protein) permeable to molecules 5000 daltons or less
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Describe the intermembrane space of the mitochondria
contains enzymes that use the ATP passing out of the matrix to phosphorylate other nucleotides
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explain oxidative phosphorolation
consumes O2 and adds a phosphate group to ADP to form ATP
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