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15 Cards in this Set
- Front
- Back
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Reverse genetics |
An approach to discover the function of a gene by analyzing the phenotypic effects of specific gene sequences obtained by DNA sequencing Reverse genetics seeks to find what phenotypes arise as a result of particular genetic sequences Reverse genetics (protein → cloned gene → mutant) |
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Forward genetics |
Employs the use of mutagens to find the genetic basis of the phenotype or trait that would be observed by genetic mapping Genetically modified cells are studied to learn about the function of the gene or protein Forward genetics (mutant → cloned gene → protein) |
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Huntington’s disease (Dominant mutation) |
Defective neural protein (huntingtin) may assemble into aggregates Aggregates cause damage to neural tissue Autosomal dominant (highlighted portion) One gene is mutated (AHD), other gene is functional (A+ )AHD/A+ still results in Huntington’s Disease = dominant mutatio |
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Cystic fibrosis (Recessive mutation) |
Defective chloride channel (CFTR) in epithelial cells leads to excessive mucus in lungs Autosomal recessive (highlighted portion )Both genes need to be mutated (ACFTR) to cause disease = recessive mutation |
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Genetic Suppression |
Arises when a second mutation (suppressor mutation) alleviates or reverts the phenotypic effects of an already existing mutation |
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Synthetic lethality |
Arises when a combination of mutations in two or more genes leads to cell death, whereas a mutation in only one of these genes does not, and by itself is said to be viable |
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Palindromic sequences (know this) |
A DNA locus whose 5'-to-3' sequence is identical on each DNA strand The sequence is the same when read left to right on one strand and right to left on the other |
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EcoRI |
Enzyme derived from the bacterium Escherichia coli Cuts foreign DNA |
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Role of restriction enzymes in bacteria |
Defense mechanisms against invading viruses in bacteria Bacterial host DNA is methylated by a modification enzyme (methylase) to protect it from the restriction enzyme’s activity |
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Taq polymerase |
DNA polymerase used in PCR Heat resistant Derived from thermophilic bacterium Thermus aquaticus (lives in hot springs) |
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Ampicillin resistant (ampr) plasmid vector |
Cut with EcoR1 and cDNA ligated to plasmid Ampr recombinant plasmids and screened and allowed to grow |
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Sequencing of DNA (Old method) |
Step-wise Addition of dideoxynucleotides (ddNTP) can be used to sequence DNAddNTP is radioactively labeled and added to primer Addition of ddNTP capped primer in low concentration to template strand prevent further elongation of DNA chain by DNA polymerase Daughter strands are denatured and separated by electrophoresis Each rung on the gel corresponds to the addition on one nucleotide Allows for the DNA sequence to be determined |
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Cre-Lox recominbation |
Recombination technique used to carry out deletions, insertions, translocations and inversions at specific sites in the DNA of cells; Can be used to knockout tissues during specific points of development Used because ESC knockouts may be embryonically lethal |
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Uses siRNA to knockdown (inhibit expression of) genetic information |
siRNA = small interfering RNAPrincipleDNA information is not deleted Specific messenger RNA is targeted siRNA specific to gene of interest are generated siRNA hybridize to specific mRNA and target it for degradation The RNA expressed by the gene of interest is only degraded The gene itself on chromosomal DNA stays intact |
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Northern analysis (know this) |
Technique used in to study gene expression by detection of RNA Mechanism Purify RNA from cells or tissue Separate RNAs by agarose gel electrophoresis Transfer RNAs to membrane Hybridize to specific probe (e.g., radioactive labeled DNA) |
