Honors - Dna 1112
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The document discusses several topics related to molecular biology and DNA replication. It provides figures and diagrams to illustrate key concepts such as: 1) The DNA replication process, with leading and lagging strands synthesized in opposite directions and RNA primers used to initiate DNA synthesis. 2) Restriction enzymes cutting DNA at specific sites to generate sticky ends for recombinant DNA techniques. 3) The polymerase chain reaction amplifying a specific DNA region using primers and repeated cycles of denaturation, annealing and extension. 4) DNA sequencing methods like Sanger sequencing using labeled dideoxynucleotides to terminate DNA strands at different positions for detection.
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Honors - Dna 1112
- 1. Molecular Biology Honors Biology Edgar
- 17. DNA Replication
- 24. Exonuclease
- 27. Fig. 16-UN5
- 28. Fig. 16-13 Single-strand binding Primase proteins Topoisomerase 3 5 RNA 3 primer 5 5 3 Helicase
- 30. Fig. 16-16b6 3 5 5 3 Template strand 3 5 RNA primer 3 1 5 Okazaki 3 fragment 5 3 5 1 3 5 3 2 1 5 5 3 3 5 2 1 5 3 3 1 5 2 Overall direction of replication
- 32. Fig. 16-16a Overview Origin of replication Leading strand Lagging strand Lagging strand 2 1 Leading strand Overall directions of replication
- 33. Helicase
- 39. Fig. 20-3-1 Restriction site DNA 5 3 3 5 1 Restriction enzyme cuts sugar-phosphate backbones. Sticky end
- 40. Fig. 20-3-2 Restriction site DNA 5 3 3 5 1 Restriction enzyme cuts sugar-phosphate backbones. Sticky end 2 DNA fragment added from another molecule cut by same enzyme. Base pairing occurs. One possible combination
- 41. Fig. 20-3-3 Restriction site DNA 5 3 3 5 1 Restriction enzyme cuts sugar-phosphate backbones. Sticky end 2 DNA fragment added from another molecule cut by same enzyme. Base pairing occurs. One possible combination 3 DNA ligase seals strands. Recombinant DNA molecule
- 42. Fig. 20-9a TECHNIQUE Mixture of Power DNA mol- source ecules of – Cathode Anode + different sizes Gel 1 Power source – + Longer molecules 2 Shorter molecules
- 43. Fig. 20-9b RESULTS
- 45. Fig. 20-10 Normal -globin allele Normal Sickle-cell allele allele 175 bp 201 bp Large fragment DdeI DdeI DdeI DdeI Large fragment Sickle-cell mutant -globin allele 376 bp 376 bp Large fragment 201 bp 175 bp DdeI DdeI DdeI (a) DdeI restriction sites in normal and (b) Electrophoresis of restriction fragments sickle-cell alleles of -globin gene from normal and sickle-cell alleles
- 47. Restriction Enzyme Lab • HINTS: • Lambda DNA/PstI: • pMAP is 5615bp • You should not be • There are able to see beyond – 2 PstI sites. the 805bp band. – 1 HpaI site. • Fine the 11,490bp – 1 SspI site and the 805bp as reference.
- 72. Gene Regulation
- 74. Fig. 18-6 Signal NUCLEUS Chromatin Chromatin modification DNA Gene available for transcription Gene Transcription RNA Exon Primary transcript Intron RNA processing Tail Cap mRNA in nucleus Transport to cytoplasm CYTOPLASM mRNA in cytoplasm Translation Degradation of mRNA Polypeptide Protein processing Active protein Degradation of protein Transport to cellular destination Cellular function
- 75. Fig. 18-8-1 Poly-A signal sequence Enhancer Proximal Termination (distal control elements) control elements region Exon Intron Exon Intron Exon DNA Upstream Downstream Promoter
- 76. Fig. 18-8-2 Poly-A signal sequence Enhancer Proximal Termination (distal control elements) control elements region Exon Intron Exon Intron Exon DNA Upstream Downstream Promoter Transcription Primary RNA Exon Intron Exon Intron Exon Cleaved 3 end transcript 5 of primary transcript Poly-A signal
- 77. Fig. 18-8-3 Poly-A signal sequence Enhancer Proximal Termination (distal control elements) control elements region Exon Intron Exon Intron Exon DNA Upstream Downstream Promoter Transcription Primary RNA Exon Intron Exon Intron Exon Cleaved 3 end transcript 5 of primary RNA processing transcript Intron RNA Poly-A signal Coding segment mRNA 3 Start Stop 5 Cap 5 UTR codon codon 3 UTR Poly-A tail
- 78. Fig. 18-9-1 Activators Promoter Gene DNA Enhancer Distal control TATA element box
- 79. Fig. 18-9-2 Activators Promoter Gene DNA Enhancer Distal control TATA element box General transcription factors DNA-bending protein Group of mediator proteins
- 80. Fig. 18-9-3 Activators Promoter Gene DNA Enhancer Distal control TATA element box General transcription factors DNA-bending protein Group of mediator proteins RNA polymerase II RNA polymerase II Transcription initiation complex RNA synthesis
- 81. Fig. 18-10 Enhancer Promoter Control Albumin gene elements Crystallin gene LIVER CELL LENS CELL NUCLEUS NUCLEUS Available activators Available activators Albumin gene not expressed Albumin gene expressed Crystallin gene not expressed Crystallin gene expressed (a) Liver cell (b) Lens cell
- 82. Fig. 18-2 Precursor Feedback inhibition trpE gene Enzyme 1 trpD gene Regulation of gene expression Enzyme 2 trpC gene trpB gene Enzyme 3 trpA gene Tryptophan (a) Regulation of enzyme (b) Regulation of enzyme activity production
- 83. Fig. 18-3a trp operon Promoter Promoter Genes of operon DNA trpR trpE trpD trpC trpB trpA Regulatory Operator gene Start codon Stop codon 3 mRNA RNA mRNA 5 5 polymerase E D C B A Protein Inactive Polypeptide subunits that make up repressor enzymes for tryptophan synthesis (a) Tryptophan absent, repressor inactive, operon on
- 84. Fig. 18-3b-1 DNA No RNA made mRNA Protein Active repressor Tryptophan (corepressor) (b) Tryptophan present, repressor active, operon off
- 85. Fig. 18-3b-2 DNA No RNA made mRNA Protein Active repressor Tryptophan (corepressor) (b) Tryptophan present, repressor active, operon off
- 86. Fig. 18-4a Regulatory Promoter gene Operator DNA lacI lacZ No RNA made 3 mRNA RNA 5 polymerase Active Protein repressor (a) Lactose absent, repressor active, operon off
- 87. Fig. 18-4b lac operon DNA lacI lacZ lacY lacA RNA polymerase 3 mRNA mRNA 5 5 Protein -Galactosidase Permease Transacetylase Allolactose Inactive (inducer) repressor (b) Lactose present, repressor inactive, operon on
- 88. Fig. 18-5 Promoter Operator DNA lacI lacZ CAP-binding site RNA polymerase Active binds and cAMP CAP transcribes Inactive Inactive lac CAP repressor Allolactose (a) Lactose present, glucose scarce (cAMP level high): abundant lac mRNA synthesized Promoter Operator DNA lacI lacZ CAP-binding site RNA polymerase less likely to bind Inactive CAP Inactive lac repressor (b) Lactose present, glucose present (cAMP level low): little lac mRNA synthesized
- 89. Epigenetics
- 98. RNAi
- 103. Vascular Endothelial Growth Factor
- 104. Genetic Engineering & Cloning
- 105. Cloning
- 106. Fig. 20-4-1 TECHNIQUE Hummingbird cell Bacterial cell lacZ gene Restriction Sticky Gene of interest site ends ampR gene Bacterial Hummingbird plasmid DNA fragments
- 107. Fig. 20-4-2 TECHNIQUE Hummingbird cell Bacterial cell lacZ gene Restriction Sticky Gene of interest site ends ampR gene Bacterial Hummingbird plasmid DNA fragments Nonrecombinant plasmid Recombinant plasmids
- 108. Fig. 20-4-3 TECHNIQUE Hummingbird cell Bacterial cell lacZ gene Restriction Sticky Gene of interest site ends ampR gene Bacterial Hummingbird plasmid DNA fragments Nonrecombinant plasmid Recombinant plasmids Bacteria carrying plasmids
- 109. Fig. 20-4-4 TECHNIQUE Hummingbird cell Bacterial cell lacZ gene Restriction Sticky Gene of interest site ends ampR gene Bacterial Hummingbird plasmid DNA fragments Nonrecombinant plasmid Recombinant plasmids Bacteria carrying plasmids RESULTS Colony carrying non- Colony carrying recombinant recombinant plasmid plasmid with disrupted lacZ gene with intact lacZ gene One of many bacterial clones
- 111. mtDNA Theories, Molecular Basis and Real-World Application
- 114. “The Other Genome” mtDNA
- 115. Endosymbiotic Theory
- 123. DNA Laboratory at Milton Academy • Isolate DNA from cheek cells. • Polymerase Chair Reaction • Electrophoresis • Sequence DNA
- 124. mtDNA Control Region
- 129. Taq DNA Polymerase
- 130. Fig. 20-8a 5 3 TECHNIQUE Target sequence Genomic DNA 3 5
- 131. Fig. 20-8b 1 Denaturation 5 3 3 5 2 Annealing Cycle 1 yields Primers 2 molecules 3 Extension New nucleo- tides
- 132. Fig. 20-8c Cycle 2 yields 4 molecules
- 133. Fig. 20-8d Cycle 3 yields 8 molecules; 2 molecules (in white boxes) match target sequence
- 135. Gel Electrophoresis
- 136. DNA Sequencing
- 137. Chain Termination Methods Sanger Methods
- 139. Fig. 20-12 TECHNIQUE DNA Primer Deoxyribonucleotides Dideoxyribonucleotides (template strand) (fluorescently tagged) dATP ddATP dCTP ddCTP dTTP ddTTP DNA polymerase dGTP ddGTP DNA (template Labeled strands strand) Shortest Longest Direction of movement Longest labeled strand of strands Detector Laser Shortest labeled strand RESULTS Last base of longest labeled strand Last base of shortest labeled strand
- 140. Fig. 20-12a TECHNIQUE DNA Primer Deoxyribonucleotides Dideoxyribonucleotides (template strand) (fluorescently tagged) dATP ddATP dCTP ddCTP DNA dTTP ddTTP polymerase dGTP ddGTP
- 141. Fig. 20-12b TECHNIQUE DNA (template Labeled strands strand) Shortest Longest Direction of movement Longest labeled strand of strands Detector Laser Shortest labeled strand RESULTS Last base of longest labeled strand Last base of shortest labeled strand
- 142. Trace File
- 143. Amplification and clonal selection
- 145. High-throughput sequencing Next-Gen Sequencing