pre-midterm content
Genetic Approach to Studying Mutants
Information gained from genetic studies of mutants can confirm amino acid functionalities and elucidate roles in biological processes.
Permissive Conditions
In temperature-sensitive mutants, permissive conditions allow the mutated gene product to behave normally, obscuring the mutant phenotype.
Roles of Model Organisms
Model organisms enable comprehensive insights into biological systems on cellular, tissue, organ, and systemic levels.
Functions of DNA
Replication: Must replicate faithfully for proper growth and cell division.
Information Storage: Saves instructions for protein synthesis and essential functions.
Mutation: Allows for genetic variation through mutations, fostering natural selection and evolution.
Roles of Bonds in DNA Structure
Phospho-diester Bonds: Stabilize the structure by linking phosphate and sugar in the backbone.
Hydrogen Bonds: Connect base pairs between DNA strands, essential for double helix stability.
Hydrophobic Interactions: Affect DNA’s interaction with water, influencing molecular structure.
Non-Covalent Interactions
Interactions maintaining double-helical conformation include ionic, hydrogen, van der Waals, and hydrophobic interactions.
Meselson-Stahl Experiment
Demonstrated semiconservative DNA replication using E. coli grown in N-15, transferring to N-14, and analyzing DNA density after centrifugation. Each daughter molecule contains one old and one new strand.
Polarity of DNA Strands
DNA strands are antiparallel, with opposing 5’ and 3’ ends, and are complementary, meaning the sequence of one strand dictates the other.
DNA Composition Ratios
If C content is 20%, then G also is 20%; thus, AT content is 60%, leading to T content of 30%.
Nucleoside vs Nucleotide
Nucleosides consist of sugar and base; nucleotides include sugar, base, and phosphate. dNTP stands for deoxynucleotide triphosphate.
High Salt Concentration and Denaturation
High salt stabilizes negatively charged phosphates in DNA, reducing strand repulsion, resulting in higher melting temperatures due to duplex stability.
Classes of DNA Sequences
Highly Repetitive: Fast renaturation.
Moderately Repetitive: Slower renaturation.
Unique: Slowest renaturation.
Cot Analysis
Measures genomic DNA complexity through DNA reassociation kinetics.
Absorbance Comparison
Single-stranded DNA will show a higher concentration than double-stranded DNA at the same absorbance (260 nm).
Bacterial vs Eukaryotic Genome Complexity
Bacterial genomes are primarily unique without repetitive sequences, while eukaryotic genomes feature high and moderate repeat sequences and unique sequences with varying renaturation kinetics.
C-value Paradox
There is no correlation between genome size (C-value) and organismal complexity.
Factors Influencing DNA Renaturation
High salt, low temperature, DNA concentration, fragment size, and overall genomic complexity.
New Species DNA Complexity Evaluation
a) 3 classes found: 25% highly repetitive, 25% moderately repetitive, 50% unique.
Prokaryotic Topoisomerase Distinctions
Topoisomerase I introduces positive supercoiling; Gyrase introduces negative supercoiling; Topoisomerase cuts single strands, while Gyrase cuts double strands and requires ATP.
Topological Isomers of DNA
Variants of DNA differing in their supercoiling state.
Importance of DNA Supercoiling
Ensures compact DNA packaging for cell survival, facilitates transcription, protects against damage, and provides energy for unwinding during replication/transcription.
DNA vs RNA Structural Differences
Primary: Uracil (RNA) replaces thymine (DNA).
Secondary: DNA: double helix; RNA: folds into stem-loops and hairpins.
Circular DNA Twisting and Writhe Calculation
4800 bp circular DNA with L=450 results in 480 twist and -30 writhe (negatively supercoiled).
Mutation Observation in Histones
Histones are conserved due to their essential, stable role in chromatin structure.
Eukaryotic Chromosomes Features
Linear DNA with nucleosomes (histone structures) wound into higher-order folds, with centromeres for spindle attachment and telomeres for chromosome end protection.
Histone Amino Acid Composition
Histones are rich in lysine and arginine, stabilizing DNA’s negative charge and facilitating its binding.
Non-Histone Proteins in Chromatin
Proteins such as MARs and SMC that assist in structural organization and transcription regulation within chromatin.
Heterochromatin vs Euchromatin
Heterochromatin: tightly packed, transcriptionally inactive; Euchromatin: loosely packed, active in transcription and replication.
Histones per Kilobase Comparison
More histones are expected in heterochromatin due to its tightly packed structure requiring additional binding proteins.
Gene Expression and Chromatin State
Genes in euchromatin are expressed in active cell types; genes in heterochromatin are usually not expressed.
Developmental Chromatin Changes
Genes that become euchromatin in later development allow necessary muscle expression.
Human Genomic Measurements
a) Diploid genome: 6 billion bp = 6 million kb.
b) Average chromosome length ~4.43 cm if in relaxed B-DNA.
c) ~13 million complete turns per chromosome.
d) Average of 1304-1739 genes per chromosome.
33-36. Homologous vs Non-Homologous Chromosomes- Homologous chromosomes: similar pairs from each parent.- Non-Homologous chromosomes: distinct and unrelated pairs.- Both genders have 23 homologous chromosomes.
Cell Division Purpose
Eukaryotes: growth, repair, reproduction.
Prokaryotes: reproduction.
DNA Replication vs Cell Division
DNA replication is the copying of genetic material, whereas cell division is the actual division into daughter cells.
Mitosis vs Meiosis Purpose
Mitosis: tissue growth and repair.
Meiosis: production of gametes, promoting genetic variability.
Genetic Variability
Results from crossing over in prophase I of meiosis.
Cell Cycle Definition and Stages
A sequence of events leading to cell division; includes interphase and mitosis.
Cell Cycle Checkpoints
G1: checks cell size/environment; S phase: replication progress; G2: checks DNA replication; M: ensures chromosome alignment.
Triggers for Cell Cycle Arrest
DNA damage, improper mass, defective replication, lack of nutrients.
Chromosome Segregation
Mitosis occurs in anaphase; meiosis occurs in meiosis I anaphase.
Chromatid Segregation
Mitosis occurs in anaphase; meiosis occurs in meiosis II anaphase.