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Gene Regulation Ch. 18 Figure 18.2 Precursor Feedback inhibition trpE gene Enzyme 1 trpD gene Enzyme 2 Regulation of gene expression trpC gene trpB gene Enzyme 3 trpA gene Tryptophan (a) Regulation of enzyme activity (b) Regulation of enzyme production Figure 18.3a trp operon Promoter Promoter Genes of operon DNA trpR Regulatory gene mRNA trpE 3 Operator RNA Start codon polymerase mRNA 5 trpD trpC trpB trpA C B A Stop codon 5 E Protein Inactive repressor D Polypeptide subunits that make up enzymes for tryptophan synthesis (a) Tryptophan absent, repressor inactive, operon on Figure 18.3b-1 DNA mRNA Protein Active repressor Tryptophan (corepressor) (b) Tryptophan present, repressor active, operon off Figure 18.3b-2 DNA No RNA made mRNA Protein Active repressor Tryptophan (corepressor) (b) Tryptophan present, repressor active, operon off Figure 18.3 trp operon Promoter Promoter Genes of operon DNA trpE trpR trpD trpC trpB trpA C B A Operator Regulatory gene 3 RNA polymerase Start codon Stop codon mRNA 5 mRNA 5 E Protein Inactive repressor D Polypeptide subunits that make up enzymes for tryptophan synthesis (a) Tryptophan absent, repressor inactive, operon on DNA No RNA made mRNA Protein Active repressor Tryptophan (corepressor) (b) Tryptophan present, repressor active, operon off Figure 18.4a Regulatory gene DNA Promoter Operator lacI lacZ No RNA made 3 mRNA 5 Protein RNA polymerase Active repressor (a) Lactose absent, repressor active, operon off Figure 18.4b lac operon lacI DNA lacZ lacY lacA Permease Transacetylase RNA polymerase 3 mRNA 5 mRNA 5 -Galactosidase Protein Allolactose (inducer) Inactive repressor (b) Lactose present, repressor inactive, operon on Figure 18.4 Regulatory Promoter gene DNA Operator lacI lacZ No RNA made 3 mRNA RNA polymerase 5 Active repressor Protein (a) Lactose absent, repressor active, operon off lac operon DNA lacI lacZ lacY lacA RNA polymerase 3 mRNA 5 mRNA 5 -Galactosidase Protein Allolactose (inducer) Inactive repressor (b) Lactose present, repressor inactive, operon on Permease Transacetylase Figure 18.7 Histone tails Amino acids available for chemical modification DNA double helix Nucleosome (end view) (a) Histone tails protrude outward from a nucleosome Acetylated histones Unacetylated histones (b) Acetylation of histone tails promotes loose chromatin structure that permits transcription Figure 18.18-1 Nucleus Embryonic precursor cell Master regulatory gene myoD Other muscle-specific genes DNA OFF OFF Figure 18.18-2 Nucleus Embryonic precursor cell Myoblast (determined) Master regulatory gene myoD Other muscle-specific genes DNA OFF OFF mRNA OFF MyoD protein (transcription factor) Figure 18.18-3 Nucleus Embryonic precursor cell Master regulatory gene myoD Other muscle-specific genes DNA Myoblast (determined) OFF OFF mRNA OFF MyoD protein (transcription factor) mRNA MyoD Part of a muscle fiber (fully differentiated cell) mRNA Another transcription factor mRNA mRNA Myosin, other muscle proteins, and cell cycle– blocking proteins Figure 18.19a Head Thorax Abdomen 0.5 mm Dorsal BODY AXES Anterior Left Ventral (a) Adult Right Posterior Figure 18.19b Follicle cell 1 Egg Nucleus developing within ovarian follicle Egg Nurse cell 2 Unfertilized egg Depleted nurse cells Egg shell Fertilization Laying of egg 3 Fertilized egg Embryonic development 4 Segmented embryo Body segments 0.1 mm Hatching 5 Larval stage (b) Development from egg to larva Figure 18.22 100 m RESULTS Anterior end Fertilization, translation of bicoid mRNA Bicoid mRNA in mature unfertilized egg Bicoid mRNA in mature unfertilized egg Bicoid protein in early embryo Bicoid protein in early embryo LECTURE PRESENTATIONS For CAMPBELL BIOLOGY, NINTH EDITION Jane B. Reece, Lisa A. Urry, Michael L. Cain, Steven A. Wasserman, Peter V. Minorsky, Robert B. Jackson Chapter 20 Biotechnology Lectures by Erin Barley Kathleen Fitzpatrick © 2011 Pearson Education, Inc. Figure 20.2 Bacterium 1 Gene inserted into plasmid Bacterial Plasmid chromosome Recombinant DNA (plasmid) Cell containing gene of interest Gene of interest 2 Plasmid put into bacterial cell DNA of chromosome (“foreign” DNA) Recombinant bacterium 3 Host cell grown in culture to form a clone of cells containing the “cloned” gene of interest Protein expressed from gene of interest Gene of interest Protein harvested Copies of gene Basic research on gene 4 Basic research and various applications Basic research on protein Gene for pest Gene used to alter Protein dissolves Human growth resistance inserted bacteria for cleaning blood clots in heart hormone treats into plants up toxic waste attack therapy stunted growth Figure 20.2a Bacterium 1 Gene inserted into plasmid Bacterial Plasmid chromosome Recombinant DNA (plasmid) Recombinant bacterium Gene of interest 2 Plasmid put into bacterial cell Cell containing gene of interest DNA of chromosome (“foreign” DNA) Figure 20.2b 3 Host cell grown in culture to form a clone of cells containing the “cloned” gene of interest Protein expressed from gene of interest Gene of interest Protein harvested Copies of gene Basic research on gene 4 Basic research and various applications Basic research on protein Gene for pest Gene used to alter Protein dissolves Human growth resistance inserted bacteria for cleaning blood clots in heart hormone treats into plants up toxic waste attack therapy stunted growth Figure 20.3-1 Restriction site 5 GAATTC CTTAAG DNA 3 5 3 1 Restriction enzyme cuts sugar-phosphate backbones. 5 3 3 5 5 Sticky 3 end 3 5 Figure 20.3-2 Restriction site 5 3 GAATTC CTTAAG DNA 5 3 1 Restriction enzyme cuts sugar-phosphate backbones. 5 5 3 3 5 Sticky 3 3 5 end 5 2 DNA fragment added 3 3 5 from another molecule cut by same enzyme. Base pairing occurs. 5 3 3 5 3 5 G AATT C C TTAA G G AATT C C TTAA G 53 5 3 One possible combination 3 5 Figure 20.3-3 Restriction site 5 3 GAATTC CTTAAG DNA 5 3 1 Restriction enzyme cuts sugar-phosphate backbones. 5 3 5 3 5 Sticky 3 3 5 end 5 2 DNA fragment added 3 3 5 from another molecule cut by same enzyme. Base pairing occurs. 5 3 5 3 3 DNA ligase 3 5 G AATT C C TTAA G G AATT C C TTAA G 53 5 3 3 5 One possible combination seals strands 5 3 3 Recombinant DNA molecule 5 Figure 20.4 TECHNIQUE Bacterial plasmid R amp gene Hummingbird cell lacZ gene Restriction site Sticky ends Gene of interest Hummingbird DNA fragments Recombinant plasmids Nonrecombinant plasmid Bacteria carrying plasmids RESULTS Colony carrying nonrecombinant plasmid with intact lacZ gene Colony carrying recombinant plasmid with disrupted lacZ gene One of many bacterial clones Figure 20.4a-1 TECHNIQUE Bacterial plasmid ampR gene Hummingbird cell lacZ gene Restriction site Sticky ends Gene of interest Hummingbird DNA fragments Figure 20.4a-2 TECHNIQUE Bacterial plasmid ampR gene Hummingbird cell lacZ gene Restriction site Sticky ends Gene of interest Hummingbird DNA fragments Recombinant plasmids Nonrecombinant plasmid Figure 20.4a-3 TECHNIQUE Bacterial plasmid ampR gene Hummingbird cell lacZ gene Restriction site Sticky ends Gene of interest Hummingbird DNA fragments Recombinant plasmids Nonrecombinant plasmid Bacteria carrying plasmids Figure 20.4b Bacteria carrying plasmids RESULTS Colony carrying nonrecombinant plasmid with intact lacZ gene Colony carrying recombinant plasmid with disrupted lacZ gene One of many bacterial clones Figure 20.8a 5 TECHNIQUE 3 Target sequence Genomic DNA 3 5 Figure 20.8b 1 Denaturation 5 3 3 5 2 Annealing Cycle 1 yields 2 molecules Primers 3 Extension New nucleotides Figure 20.8c Cycle 2 yields 4 molecules Figure 20.8d Cycle 3 yields 8 molecules; 2 molecules (in white boxes) match target sequence Figure 20.8 5 TECHNIQUE 3 Target sequence Genomic DNA 1 Denaturation 3 5 5 3 3 5 2 Annealing Cycle 1 yields 2 molecules Primers 3 Extension New nucleotides Cycle 2 yields 4 molecules Cycle 3 yields 8 molecules; 2 molecules (in white boxes) match target sequence Figure 20.9a TECHNIQUE 1 Mixture of DNA molecules of different sizes Power source Cathode Anode Wells Gel 2 Power source Longer molecules Shorter molecules Figure 20.9b RESULTS Figure 20.10b Normal Sickle-cell allele allele Large fragment 376 bp 201 bp 175 bp (b) Electrophoresis of restriction fragments from normal and sickle-cell alleles Figure 20.19a TECHNIQUE Mammary cell donor Egg cell donor 1 Egg cell from ovary Cultured mammary cells 2 Nucleus removed 3 Cells fused Nucleus from mammary cell Figure 20.19b Nucleus from mammary cell 4 Grown in culture Early embryo 5 Implanted in uterus of a third sheep Surrogate mother 6 Embryonic development RESULTS Lamb (“Dolly”) genetically identical to mammary cell donor Figure 20.23 Cloned gene 1 Insert RNA version of normal allele into retrovirus. Viral RNA Retrovirus capsid 2 Let retrovirus infect bone marrow cells that have been removed from the patient and cultured. 3 Viral DNA carrying the normal allele inserts into chromosome. Bone marrow cell from patient 4 Inject engineered cells into patient. Bone marrow Figure 20.26 TECHNIQUE Agrobacterium tumefaciens Ti plasmid Site where restriction enzyme cuts T DNA DNA with the gene of interest RESULTS Recombinant Ti plasmid Plant with new trait
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