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Making a mutant DNA Deletion: Deletes a hunk of DNA Insertion: Inserts a hunk of DNA Site-directed: Modifies a specific nucleotide Random: Introduces random changes in the DNA Mutagenesis is used to alter the DNA structure (sequence) in a known way by either deleting nucleotides, inserting nucleotides, or changing a single nucleotide at a defined location Random mutagenesis of DNA may be performed over the whole piece of DNA by exposing the DNA to chemicals (mutagens) that react with the DNA and change the specificity for base pairing or by using oligonucleotides that contain random, deliberate mistakes in the sequence Mutants are then selected or screened for changes in function of the protein or RNA product This technique allows you to define specific amino acids that are essential to the function of the protein and to determine which amino acids can be replaced by which other amino acids and still conserve function In deletion or insertion mutagenesis, restriction enzymes are used to generate DNA with a specific fragment missing or with another piece of DNA inserted This change in the DNA sequence can then be used to produce RNA or protein containing a deletion or insertion of amino acids in the protein This is useful in determining the gross features of the gene structure that are necessary to preserve a functional gene or to express a functional protein product For example, a signal sequence that directs the synthesized protein to the mitochondrial matrix can be placed in the sequence of a protein that is normally cytosolic This mutant protein will be expressed as a mitochondrial matrix protein With site-directed mutagenesis, a change in the DNA sequence can be introduced at any specific site An oligonucleotide is annealed to a single-strand copy of the DNA that you want to mutate This oligonucleotide contains the correct (complementary) base at every position except the one you want to change At the mutated position, there is a mismatch After the oligonucleotide is annealed at the proper position, the DNA is fully replicated using DNA polymerase and then sealed with ligase When the vector is introduced into the host cell and replicated during cell division, some of the progeny cells will get DNA that has used the mutant strand as the template for DNA replication There are clever ways to increase your chances of getting only cells containing the mutant DNA These involve selectively destroying the wild-type (nonmutated).

The older your child gets, the less likely she is to stay in one place for long Her curiosity takes her around corners, up stairs, and into new rooms At the same time, as a parent you are becoming increasingly secure that she need not be

strand Site-directed mutagenesis is used to change single amino acids in proteins or single bases in RNA or DNA The technique has been very useful in determining the function of a specific amino acid residue in enzyme catalysis, binding of a ligand, or stabilizing a protein It has also been possible to selectively change the activity and specificity of some enzymes using this technique (Fig 6-6)

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under your watchful eye absolutely every moment So your child enjoys a new freedom to explore the nooks and crannies of her world What many of these new places have in common is that your child no longer can see you, and you can no longer see her But that doesn t mean she doesn t still want to tell you things Putting yourself in your child s place, you can quickly see the problem As any person with a hearing impairment can tell you, the usefulness of sign language disappears when folks aren t face to face But sounds, on the other hand, can be heard even shouted from room to room So, as signers start to move farther and farther afield, learning the words behind the signs takes on an urgency not felt when everyone could be counted on to stay in one place

Venera probes, 129 Venus, 123 125, 128 132, 139 144, 460 vernal equinox, 17 Virgo, 34, 68 virtual reality, 480 visible light, 450 visual magnitude, 269 Volans, 66 Voyager, 174, 176, 182, 184, 185, 188 waning crescent moon, 91 93 waning gibbous moon, 91 93 War of the Worlds, 154 wavelength, 449 waxing crescent moon, 90 93 waxing gibbous moon, 91 93 Weather Underground, 15, 27, 61, 181, 186, 503 Wells, H G, 154 western hemisphere, 8 Whipple, Fred, 264 white dwarf, 324, 326 white hole, 376 white supergiant, 327 winter solstice, 18 x-ray stars, 471 x-ray telescope, 470 x-rays, 450, 469 472 yellow supergiant, 330 zenith, 9, 54 zero-g, 490 Zeus, 35, 82

Figure 6-6

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