© 2012 Pearson Education, Inc. Lecture by Edward J. Zalisko PowerPoint Lectures for Campbell Biology: Concepts & Connections, Seventh Edition Reece, Taylor, Simon, and Dickey Chapter 12 DNA Technology and Genomics

Figure 12.1A Animation: Cloning a Gene

Figure 12.1B_s1 E. coli bacterium Bacterial chromosome A plasmid is isolated. Gene of interest Plasmid The cell’s DNA is isolated. DNA A cell with DNA containing the gene of interest 1 2

Figure 12.1B_s2 E. coli bacterium Bacterial chromosome A plasmid is isolated. Gene of interest Plasmid The cell’s DNA is isolated. DNA A cell with DNA containing the gene of interest The plasmid is cut with an enzyme. The cell’s DNA is cut with the same enzyme. Gene of interest

Figure 12.1B_s3 E. coli bacterium Bacterial chromosome A plasmid is isolated. Gene of interest Plasmid The cell’s DNA is isolated. DNA A cell with DNA containing the gene of interest The plasmid is cut with an enzyme. The cell’s DNA is cut with the same enzyme. Gene of interest The targeted fragment and plasmid DNA are combined.

Figure 12.1B_s4 E. coli bacterium Bacterial chromosome A plasmid is isolated. Gene of interest Plasmid The cell’s DNA is isolated. DNA A cell with DNA containing the gene of interest The plasmid is cut with an enzyme. The cell’s DNA is cut with the same enzyme. Gene of interest The targeted fragment and plasmid DNA are combined. DNA ligase is added, which joins the two DNA molecules. Gene of interest Recombinant DNA plasmid

Figure 12.1B_s5 Gene of interest The recombinant plasmid is taken up by a bacterium through transformation. Recombinant bacterium Recombinant DNA plasmid 7

Figure 12.1B_s6 Gene of interest The recombinant plasmid is taken up by a bacterium through transformation. The bacterium reproduces. Clone of cells Recombinant bacterium Recombinant DNA plasmid 7 8

Figure 12.1B_s7 Gene of interest The recombinant plasmid is taken up by a bacterium through transformation. Harvested proteins may be used directly. The bacterium reproduces. Clone of cells Recombinant bacterium Recombinant DNA plasmid Genes may be inserted into other organisms

12.2 Enzymes are used to “cut and paste” DNA  Restriction enzymes cut DNA at specific sequences. –Each enzyme binds to DNA at a different restriction site.  DNA ligase joins DNA fragments together. © 2012 Pearson Education, Inc. Animation: Restriction Enzymes

Figure 12.2_s1 A restriction enzyme cuts the DNA into fragments. Restriction enzyme recognition sequence Restriction enzyme Sticky end Sticky end DNA 1 2

Figure 12.2_s2 A restriction enzyme cuts the DNA into fragments. Restriction enzyme recognition sequence Restriction enzyme Gene of interest A DNA fragment from another source is added. Sticky end Sticky end DNA 1 2 3

Figure 12.2_s3 A restriction enzyme cuts the DNA into fragments. Restriction enzyme recognition sequence Restriction enzyme Gene of interest A DNA fragment from another source is added. Two (or more) fragments stick together by base pairing. Sticky end Sticky end DNA

Figure 12.2_s4 A restriction enzyme cuts the DNA into fragments. Restriction enzyme recognition sequence Restriction enzyme Gene of interest A DNA fragment from another source is added. Two (or more) fragments stick together by base pairing. Sticky end Sticky end DNA ligase pastes the strands together. Recombinant DNA molecule DNA

Figure 18.3 Vectors for Carrying Recombinant DNA into Cells

Figure Expression of Transgene in Host Cell Produces Large Amounts of its Protein Product

Table 12.6

Figure 18.5 Green Fluorescent Protein as a Reporter

Figure 12.4 C ELL NUCLEUS DNA of a eukaryotic gene RNA transcript mRNA T EST T UBE Reverse transcriptase cDNA strand being synthesized Direction of synthesis Breakdown of RNA Synthesis of second DNA strand Isolation of mRNA from the cell and the addition of reverse transcriptase; synthesis of a DNA strand cDNA of gene (no introns) Exon Intron Transcription RNA splicing (removes introns and joins exons)

Figure Transgenic Rice Rich in  -Carotene

Figure 12.8A_s3 Restriction site The gene is inserted into the plasmid. The recombinant plasmid is introduced into a plant cell. The plant cell grows into a plant. DNA carrying the new gene A plant with the new trait Recombinant Ti plasmid Plant cell DNA containing the gene for a desired trait Ti plasmid Agrobacterium tumefaciens