Gre Bio Chp 11 Biotechnology

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Restriction Enzymes

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Enzymes that recognize specific DNA sequences and cleave them. Usually cut at palendromic DNA sequences

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Sticky Ends

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Overhanging ends of DNA after enzymes cut a sequence asymmetrically. Want to bond with complementary bases. Permits manufacture of recombinant DNA

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Blunt Ends

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Enzymes cut a sequence and do not leave overhanging ends of DNA

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Recombinant DNA

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Combination of genes from different organisms

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Transformation

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Cleaved plasmids in close proximity to cleaved foreign DNA will take up the foreign DNA. Introduces small pieces of new DNA into bacteria

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Recombinant Plasmids

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Plasmids with foreign DNA integrated. Are considered DNA vectors because they are a means by which new DNA can be introduced into the bacterial cells. Can be copied and exchanged between cells.

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Functions of Transferring of DNA using Plasmids

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Figure out the function of an unknown gene, Produce clinically useful substances, Give bacteria new functions

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Selection

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Since only 1% of bacteria in a medium pick up the plasmid, antibiotic resistant genes are added to the plasmids to select out the ones who don't pick up the plasmid

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Screening

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Genes that produce a pigmented protein are put in the plasmid to determine which bacteria pick up the plasmid by which colonies are colored

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Oligonucleotides

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short sequences of DNA

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Sanger Method

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Method for sequencing DNA. Template DNA is put in 4 different test tubes with DNA polymerase, nucleotide bases, and dideoxynucleotides. Each tube has either ddATPs, ddCTPs, ddGTPs, or ddTTPs. Sequencing stops when one of these is put into the chain

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Nucleic Acid Hybridization

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Used to probe DNA for certain sequences in a Southern Blot. Cut up DNA is separated and washed in a solution containing radioactive probes which bind to complimentary DNA sequences. This segments then glow

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Radioactive Probe

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A piece of DNA with radioactive nucleotides that is complimentary in sequence to the DNA sequence you're searching for

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Restriction Fragment Length Polymorphisms (RFLP)

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Differences in the length of the fragments made by restriction enzyme digestion of two samples of DNA

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Polymerase Chain Reaction (PCR)

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Method to rapidly amplify a piece of DNA. Does not need a living organism. A test tube containing a DNA sequence, DNA polymerase, primers, and free nucleotides is heated and cooled to let the primers hydrogen bond and complimentary stands are formed

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Transgenic Animals

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May be used to produce proteins needed for human therapies by injecting DNA directly into fertilized eggs using tissue-specific promoters

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Cloning

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The diploid nucleus from a donor cell is implanted into an unfertilized egg with its nucleus removed

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cDNA

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A gene without the introns.. Contained in bacterial cells in genomic libraries. Used by researchers who want to work with one particular gene

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Northern Blotting

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Same technique as in Southern Blotting but to spot mRNA. Used to assess differential gene expression.

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Reporter Gene

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Genes scientists fuse into the regulatory region of a gene, Produce copious amounts of easily measured protein. Allow scientists to monitor levels of gene expression in a cell.