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Frameshift Mutations

Frameshift Mutations

Caused by agents like acridine that alter the reading frame of DNA

detected via gel electrophoresis

Point Mutations

Induced by mutagens like 5BU

includes missense and nonsense mutations affecting protein structure/function

involves keto-enol tautomerization

DNA Polymerase Proofreading

Uses exonuclease activity to correct mismatches during DNA replication

Mismatch Repair

Involves recognition and repair by enzymes like MutS and MutH in bacteria, distinguishes new strands from template strands by methylation patterns

Prokaryotic IS Elements and Transposons

Simple insertion sequences vs. composite and noncomposite transposons, mechanisms include replicative and conservative transposition

Eukaryotic Transposable Elements

Includes retrotransposons that use RNA intermediates, similar structure and function to prokaryotic elements

Experimental Design

Identifying suppressor mutations using mutagens to study gene-protein interactions (e.g.

Mutation Rates

Calculating mutation frequency

Cloning Process

Steps for creating DNA clones and genomic libraries, involves selection of appropriate vectors like plasmids, BACS, YACS

cDNA Libraries

Different from genomic libraries, used to study mRNA and protein expression

Gel Electrophoresis

Separates DNA/proteins by size and charge, interprets migration patterns

Southern Blotting

Denatures DNA for hybridization to detect specific sequences

PCR

Amplifies DNA segments, efficiency calculations for multiple cycles

Genome Sequencing Strategies

Includes whole-genome shotgun sequencing and coverage importance for accuracy

Annotation and Analysis

Identifies genes, SNPs, and functional regions

Comparative Genomics

Analyzes conserved domains and homologous genes

Transcriptomics and Proteomics

Techniques to study RNA and protein abundance compares gene expression across stages or conditions

Expression Vectors

Features required for expressing eukaryotic genes in prokaryotes and mammalian systems

Insulin Production

Advantages of using cDNA over genomic DNA for protein production in bacteria