DNA replication, the genetic code, gene expression, and protein synthesis
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Yeast
Both diploid and haploid life stages. Haploid and diploid cells interconvert between the two types by mating and sporulation, respectively.
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Random mutagenesis
Altering the properties of enzymes to create a “library” containing thousands of versions of the genes.
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Error-prone PCR.
“sloppy” version of PCR, in which the polymerase has a fairly high error rate (up to 2%), to amplify the wild-type sequence. Size of the library is limited by the efficiency of the cloning step
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Rolling circle error-prone PCR
first cloned into a plasmid, then the whole plasmid is amplified under error-prone conditions. This eliminates the ligation step .
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Mutator strains
the wild-type sequence is cloned into a plasmid and transformed into a mutator strain, such as Stratagene’s XL1-Red.
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Temporary mutator strains.
Temporary mutator strains can be built by over-expressing a mutator allele such as mutD5 (a dominant negative version of mutD) which limits the cell’s ability to repair DNA lesions
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Insertion mutagenesis
Finnzymes have a kit that uses a transposon-based system to randomly insert a 15-base pair sequence throughout a sequence of interest, be it an isolated insert or plasmid.
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Ethyl methanesulfonate
Another chemical mutagen. It acts by de-aminating adenine and cytosine residues causing transversion point mutations
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DNA Shuffling
A very powerful method in which members of a library (i.e. copies of same gene each with different types of mutation) are randomly shuffled.
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Caenorhabditis elegans
fr. ee-living, transparent nematode. a simple model for multi-cellular organisms.
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Connectome
comprehensive map of neural connections in the brain, and may be thought of as its "wiring diagram".
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Drosophila melanogaster
Easy to breed and because of their small size it is possible to perform experiments on 1000s of individuals.. intrinsic and extrinsic factors controlling lens cell specification and differentiation.
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Zebrafish Danio rerio
valuable model for studying human genetics and disease. Can withstand higher doses of mutagens allowing the creation of more mutants.Their cells and body parts can regenerate.
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Mus musculus
Similar genome structure and organisation and similar number of genes. Amenable to genetic manipulation..
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3R's
Replacement (avoid use of animals), Reduction (minimise number of animals) Refinement (minimise suffering)
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Reverse genetics
understand the function of a gene by analyzing the phenotypic effects of specific engineered gene sequences.
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Mutagenesis
laboratory is an important technique whereby DNA mutations are deliberately engineered to produce mutant genes, proteins, strains of bacteria
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Transgene
gene or genetic material that has been transferred naturally, or by any of a number of genetic engineering techniques from one organism to another.
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Other cards in this set
Card 2
Front
Yeast
Back
Both diploid and haploid life stages. Haploid and diploid cells interconvert between the two types by mating and sporulation, respectively.
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