Dan-Genetics Exam 3
Terms
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- Prokaryotic Gene Expression
- Lac operon Ara operon Trp operon-attenuation Heat Shock Genes DNase-footprinting Assay
- Allosteric Proteins
- Lac repressor AraC Trp repressor Steroid Hormone
- Prokaryotic Gene Transfer Requirements
- Transformation, Conjugation, or Transduction
- Types of Eukaryotic Gene Expression
- Myc-Max, Yeast Gal system, Locus Control region, Silencing
- Types of Repression
- Competition and Quenching
- Post-Translational Eukaryotic regulation
- Micro-RNAs-formation of RISC(RNA inducing silencing complex)-prevents translation RNAsplicing-Sxl gene produces Sxl protein which regulates splicing in males and females Protein Synthesis-miRISC-break mRNA Protein Modifications-Phosphorylation-addition of Epinephrin
- Lac Operon Promoter - 3 important sequences:
- CRP-cAMP binding site TATA Box Operator Binding Site
- Basal (Protein) Transcription Factors
- RNA Poly II TAF (2) - association factors TBP - TATA binding protein Transcription moves faster when Basal Factors and Activator protein bind at same time
- Eukaryotes - 3 RNA Polymerases
- RNA Pol I - rRNA (ribosomal) RNA Pol II - Protein coding RNA Pol III - tRNA and other small RNAs
- Telomere Duplication
- Telomerase binds at specific sequence Elongation New bases added to chrom. ends
- Retroposon Movement
- Retroposon codes for reverse transcriptase RT recognizes 5bp sequence and excises Retroposon Retroposon inserted into new sequence on genome
- Transposon Movement
- Codes for Transposase Transposase recognizes inverted sequence-excises Transposon Transposon moved to another part of genome Exonuclease widens gap Repair of gap using sister chromatid with or without Transposon
- Selection of Auxotrophs
- Auxotroph and Prototrophs in rich media Centrifuge Resuspend in media minus "histidine" and plus penicillin Most prototrophs die Transfer to 2 media-one with histidine and one without Screen for Auxotrophs
- Insertion Sequence Steps
- -Clone into Plasmid -Linearize Plasmid with RE -Transform into bacteria -Screen for Mutants
- Natural Gene Transfer
- -Donor DNA binds to receptor site of recipient cell -One donor strand is degraded, other is admitted into cell -Admitted strand pairs with homologous region -Donor strand is integrated into bacterial chromosome -After cell replication
- Conjugation of F-Plasmid
- -F pillus connects with F- cell -Pillus retracts and cells are drawn together -DNase cuts plasmid DNA at target site -Single strand moves across into F- cell -F+ and F- cell synthesize newly formed strands -Cells separate to form two F+ cells
- Conjugation of Hfr Strain
- -F pillus of Hfr cell connects to F- -Single strand of F plasmid moves into F- followed by chromosomal DNA -Crossovers between homologous regions -Recipient carries F plasmid, Hfr strain, and original strain (partly diploid) -Nucleases digest DNA fragments -Cell separates and now carries recombinant genome
- Generalized Transduction
- -Phage infects host -DNA is broken up into fragments -Host DNA now bears foreign gene -Cell lyses and phages are released -Phage infects another host -Recombination occurs
- Reverse Genetics
- -Clone AmpR into plasmid -Plasmid is linearized -Transformed in bacteria -Screened for antibiotic resistance -Cells are plated on Ampicillin rich media
- DNase-Footprinting Assay
- used to determine the DNA sequence that is recognized by transcription factor
- Trp Attenuation
- Tryptophan is present-ribosome moves quickly down leader codons and 3-4 stemloop is formed (transcription stops) No Tryptophan-ribosome gets stalled at trp-codons and allows 2-3 stemloop to form (transcription)