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4 Cards in this Set
- Front
- Back
Positional Cloning
DMD CF |
-We can clone a gene by forming a self-replicating DNA vector. Restriction enzymes are used to cut out the DNA of the gene of interest. The same restriction enzyme is used to create a linear DNA molecule from a purified plasmid. We then insert the DNA into the plasmid where the two will combine and anneal using their sticky ends and DNA ligase. We can then transfect a bacterial cell with the plasmid where it will continue to self-replicate.
-Using a variation of the plus-minus screening technique, they were able to isolate several clones that were deleted in the patient. One of them would prove to contain the DMD gene (chromsome walking + southern blots) -linked to chromosome 7 by linkage analysis using RFLPs. By looking for more tightly linked RFLPs, scientists showed that the gene is contained within an ~1.5Mb region (chromosome walking + jumping) |
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Linkage map
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Linkage maps use RFLP's and related genetic markers to enable researchers to localize genes with pedigree analysis
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Physical map
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In order to then clone a localized gene, a physical map is necessary. The procedure used in model organisms, such as the nematode and yeast, is to screen a genomic library with large inserts (usually in YACs) to find, and order, a series of overlapping clones, called contigs.
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Human Genome Project
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-able to describe those genes that are involved in diseases.
-bring early detection and treatment of disease, and perhaps prevention. -help speed the task to find the genes that cause these diseases |