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cloning a segment of DNA of interest - cut vector with a restriction enzyme -add ligase to add sequence of interest -put the recombinant DNA in a bacterial cell (transformation) -Amplify the DNA- make multiple copies in a culture -Isolate the recombinant DNA -Cut with restriction enzymes |
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Carrier molecules that are used in DNA cloning, self replication DNA molecules that are used to transfer foreign DNA segments between host cells |
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-small circular DNA molecules that exist in the cytoplasm of bacterial cells |
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-origin of replication, allows for replication of plasmid DNA, a selectable marker- allows for selection for cells that have plasmid DNA |
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target for a restriction enzyme- where the insert will be inserted |
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Screening for bateria with recombinant DNA |
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pBR322 vector: adding the insert disrupts a gene that otherwise would provide tetrocyclin resistance. Cells carrying recombinant DNA die in the presence of tetracycline maintain ampR |
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adding insert dirsupts the lacZ gene. Lac Z encodes beta-galactosidase
X-Gal--> (beta gal) X + gal want the white ones, not the blue |
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contains a collection of recombinant DNA that is transcribed gene of interest -copy of a fully processed mRNA |
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Gene transfered into somatic cells of the body; single target tissue -not passed on to the next generation -doesn't last |
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-destroying of white blood cells leads to a severe immune deficiency -Bone marrow transplant is a cure- but could cause an attack on the patients tissue if not a match |
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-Genetic engineering of organisms -add new gene that is incorporated into the all of the organisms cells -Change passed on to the next generation |
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Modification in embryonic cells |
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- produces a chimera- not all cells have the same genotype -inject cells into the early embryo |
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- add gene to the male pronucleus before sygamy( where the cell is feterilized) |
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-undifferentiated cells (callus) -add in particular thing to the plant -bacterium uses a tumor (Ti) vector to introduce trans-gene |
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- can give rise to all the cell types in the adult body and cells in supportive tissues- to the whole organism |
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Present in the inner cell mass- able to give rise to all of the cell types in the body but cannot form an individual |
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multipotent (adult stem cells-somatic stem cells) |
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Present in many tissues- tissue specific |
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