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19 Cards in this Set
- Front
- Back
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Expression vector |
a vector that allows expression (txn and tln) of a cloned gene |
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Antibodies |
a protein used by the immune system to bind and neutralize foreign substances |
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In situ |
in place, in their original positions |
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Autoradiogram |
an image on an x-ray film produced by the pattern of decay emissions from a distribution of a radioactive substance - has visible dark spots |
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Hybridization |
Screening a genomic library is different from screening a cDNA library because the filter is incubated with labeled single-stranded DNA, which forms hybrids with complementary ssDNA molecules bound to the filter |
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Probes |
a fragment of DNA or RNA of variable length which can be radioactively labeled. It can then be used in DNA or RNA samples to detect presence of nucleotide sequences that are complementary sequences to it |
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Heterologous probes |
a probe made from a clones equivalent gene |
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Conserved genes |
If the species are closely related, the heterologous probe should be able to anneal to the gene of interest |
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Guessmer process/oligos |
a) If a protein has been purified and that some of the aa sequence are known, it is possible to deduce the DNA sequence from the aa sequence b) Synthetic ssDNA (oligo) can then be made, labeled, and used as a probe for library screening c) Degeneracy in the genetic code means that a mixture of oligonucleotides (guessmer) must be prepared |
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Complementation |
a) Well defined mutants may be used to clone genes by this process, in which cloned genes overcome a defect in the mutant b) clone a yeast gene this way |
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Shuttle vector |
a cloning vector capable of replicating 2 or more organisms - used in complementation of yeast gene |
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Restriction mapping |
determines the order and distances of RE cut sites in a segment of DNA |
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Gel electrophoresis |
a lab method used to separate mixture of DNA, RNA, or proteins according to molecular size - for restriction mapping |
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Subcloning |
Moving a segment of a clone to another vector (e.g. you have a clone with 2 genes in it and you want to isolate clones containing only 1 gene) |
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Southern blot |
lab technique where DNA fragments are transferred from a gel to a filter and then hybridized with a probe a) genomic DNA is cut with different RE's Useful for any form of DNA!!! |
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Northern blot |
lab technique is used for determining the size of mRNA, also show alternative splicing |
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Western blot |
lab technique determining size of proteins |
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Membrane filter? |
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Alternative splicing |
mRNA species arise from the same gene, suggesting differential use of promoters and terminators (i.e. more in brain, less in muscle) |
