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BIOCHEMISTRY 4.1 HOMEWORK NAME____________________________ USING YOUR TEXT, DEFINE THE FOLLOWING TERMS: Cloning:___________________________________________________________________________________________________ _________________________________________________________________________________________________________ Vector: __________________________________________________________________________________________________ _________________________________________________________________________________________________________ Recombinant DNA__________________________________________________________________________________________ _________________________________________________________________________________________________________ DNA Ligase: _______________________________________________________________________________________________ _________________________________________________________________________________________________________ Plasmid: __________________________________________________________________________________________________ _________________________________________________________________________________________________________ Genomics: ________________________________________________________________________________________________ _________________________________________________________________________________________________________ Complementary DNA: ______________________________________________________________________________________ _________________________________________________________________________________________________________ Polymerase Chain Reaction: _________________________________________________________________________________ _________________________________________________________________________________________________________ DNA Fingerprinting_________________________________________________________________________________________ _________________________________________________________________________________________________________ Southern Blot_____________________________________________________________________________________________ _________________________________________________________________________________________________________ DNA Microarray____________________________________________________________________________________________ _________________________________________________________________________________________________________ Transgenic________________________________________________________________________________________________ _________________________________________________________________________________________________________ 1. CLONING. When joining 2 or more DNA fragments, a researcher can adjest the sequence at the junction in a variety of subtle ways, as seen in the following exercises. a. Draw the structure of each of a linear DNA fragment produced by an EcoRI restriction digest (include those sequences remaining from the EcoRI recognition sequence). b. Draw the structures resulting from the reaction of this end sequence DNA polymerase I and four deoxynucloside triphosphases (see Fig 8-36 in the book!) 2. Selecting for recombinant plasmids When cloning a foreign DNA fragment into a plasmid, it is often useful to insert the fragment at a site that interrupts a selectable marker (such as the tetracycline-resistance gene of pBR322). The loss of function of the interrupted gene can be used to identify clones containing recombinant plasmids with foreign DNA. With a bacteriophage vector, it is not necessary to do this, yet one can easily distinguish vectors that incorporate large foreign DNA fragments from those that do not. How are these recombinant vectors identified? 3 Suggest how the researcher might use the PCR to detect the presence of the circular form of the deleted DNA in an extract from the protist.