Term
| Intro of DNA into animal cells |
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Definition
| Plasmid DNA w/ gene of interest, Ampf, ori, and gene conferring drug resistance to mammal cells. High fraction express plasmid DNA (shown by selecting for drug resistance). Some cells will permanently incorporate the plasmid into their DNA so we can study long-term gene effects. |
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Term
| Mouse as model for human devo |
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Definition
| Can study function of cloned genes in animals, not just in vitro (baby and mouse both w/ chest discoloration) |
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Term
| Production of transgenic mice |
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Definition
| Microinject plasmid into fertilized egg's pronucleus. Put eggs into mothers. About 10% of offspring will have integrated the plasmid into their DNA at RANDOM SITES, not at the site of the normal gene. |
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Term
| Intro of genes into mice via ES cells |
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Definition
| Gene is added IN ADDITION to normal gene, so we don't see what would happen if the normal gene was lost. I.e., mouse and mutant nose genes vs. just mutant nose genes. |
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Term
| Gene inactivation by homologous recombination |
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Definition
| Intro mutated gene into cell w/normal gene copy. Homologous recombination happens and mutated DNA recombines w/ normal copy. Cell will now carry mutated gene IN PLACE OF normal gene. Replaces normal gene w/ mutated inactive copy. |
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Term
| Inhibition of gene expression by antisense RNA or DNA |
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Definition
| Antisense RNA/DNA hybridizes to normal mRNA, blocking translation to protein. Blocks gene expression in cultured cells. |
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Term
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Definition
| dsRNA cleaved by Dicer into siRNA pieces (21-23 bp). RISC binds to siRNAs, unwinds siRNA into a ss molecule and complementarily pairs it to target mRNA. mRNA is cleaved where the siRNA ss bonds. Most powerful and commonly used to block gene function. |
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Term
| Direct inhibition of protein function |
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Definition
| Proteins function by interacting w/target in cell. Add antibodies, protein will be surrounded and can't interact w/target. Add inhibitory mutant protein, mutant blocks function by bonding w/target first. |
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Term
| Noncoding sequences in higher eukaryotes |
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Definition
| Introns, repetitive sequences, gene duplications and pseudogenes, intragenic DNA (DNA between genes). |
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Term
| Examples of animal viruses |
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Definition
| Retroviruses: 9 kb, RNA. Adenoviruses: 36 kb, DNA. |
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Term
| ID of introns in adenovirus mRNA |
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Definition
| When making an RNA-DNA hybrid there were these overhanging loops of DNA that weren't hybridized to RNA. Shows that the loops must be removed during RNA production. |
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