Biology Lecture 2 Genes
Examkrackers MCAT Comprehensive Course, Charles Feng
[Link] fenguin@[Link] (224) 532-0039
Genes
Definitions
o Gene: DNA that codes for a single polypeptide/mRNA/rRNA/tRNA
o Euchromatin: region of DNA containing genes being actively transcribed
o Heterochromatin: region of DNA containing genes not actively transcribed
o Genome: entire DNA sequence of an organism human: 26k-38k genes, ~1%
codes for protein
o Central Dogma: DNA (transcription)> RNA (translation)> protein
DNA
o Nucleotides: phosphate group + 5-carbon sugar + nitrogenous base
o Bases: adenine (A), guanine (G), cytosine (C), thymine (T)
o Purines: adenine & guanine, two rings
o Pyrimidines: cytosine & thymine, one ring
o Bases bound together by 5-3 phosphodiester bonds into phosphate backbone
o 53 directionality (5 phosphate, 3 OH)
o DNA strands are antiparallel, base pairing occurs between A=T,
GC - A2T C3G
o Double helix, complementary strands
Replication (DNA DNA)
Semiconservative new double stranded DNA = 1 old strand + 1
new strand
Replication begins at the origin of replication, proceeds in two
directions (bidirectional)
Continuous reading/replication can occur at leading strand, but
not at lagging strand
o Lagging strand is continuously interrupted & restarted
(Okazaki fragments)
o Typically fragments are in range of 102 - 103 bp
Page 1 of 7
Biology Lecture 2 Genes
Examkrackers MCAT Comprehensive Course, Charles Feng
[Link] fenguin@[Link] (224) 532-0039
Enzymes
o Helicase: unzips the double helix, allows primase to bind
o DNA polymerase: builds new DNA strand, can only add nucleotides to existing strand
Reads parent strand in 35 (upstream), creates new strand in 53 (downstream)
Also contains a mechanism to repair mismatched nucleotides, adds to accuracy
o Primase: creates RNA primer so DNA polymerase can start working
o DNA ligase: connects chunks of new DNA created in lagging strand
o Telomerase: lengthens telomeres (repeated sequences at ends of eukaryotic DNA, protects from being eroded)
RNA
o 2 carbon on sugar has an OH instead of an H
o Generally single stranded, though can have special secondary structures
o Has uracil (U) instead of T, so A=U and CG bonds
Types of RNA
o mRNA (messenger RNA): carries genetic info from nucleus (DNA) to cytosol
o rRNA (ribosomal RNA): forms ribosomes by combining with proteins, made in nucleolus
o tRNA (transfer RNA): carries amino acids to ribosomes
Transcription (DNA->RNA)
Initiation: initiation factors recognize promoter, cause assembly of RNA
polymerase
o Can be regulated by small changes in promoter sequence, which affect
binding
o Can be regulated by activators and repressors which bind DNA close to
promoter
o RNA polymerase unzips double helix, transcribes only
antisense (-) strand
Elongation: no proofreading, slower than replication
o RNA polymerase reads DNA in 35 direction, creates RNA
in 53 direction
Termination: RNA pol recognizes termination sequence
Prokaryotes - gene activity changes as a response to changes
in environment
Eukaryotes - gene activity changes to maintain homeostasis
Page 2 of 7
Biology Lecture 2 Genes
Examkrackers MCAT Comprehensive Course, Charles Feng
[Link] fenguin@[Link] (224) 532-0039
Operon
o Prokaryotic structure with operator, promoter, and multiple genes
o Operator: binds repressor protein to prevent RNA polymerase from binding, or binds activator protein to facilitate
RNA polymerase binding
o Promoter: binds RNA polymerase in the absence of repressor
o Multiple genes will appear on single transcript
o Lac operon: repressor protein allosterically deactivated by lactose, so increase in lactose concentration leads to
repressor not binding transcription of lactose proteins
Post-Transcriptional Processing (Eukaryotes)
o Replication primary transcript processing
o 5 cap: helps ribosome to attach, protects transcript from degradation
o poly-A tail: on 3 end, protects transcript from degradation
o Intron splicing: snRNPs (small nuclear ribonucleoproteins) cleave out introns,
leaving behind exons which are spliced together to form final transcript
o Variation in this process can make different proteins for same transcript
Translation
4 different bases, 3 base sequence codes for each amino acid 43 = 64 possibilities
Since theres only 20 amino acids, more than one sequence can code for the same
amino acid genetic code is degenerate
But each sequence can code for only one amino acid genetic code is unambiguous
3 base sequence = codon
UAA/UGA/UAG are stop codons, AUG is start codon but also codes for Met
Ribosome has a small subunit and a large subunit, produced in nucleolus
Each tRNA has an anticodon to specifically recognize codons
Page 3 of 7
Biology Lecture 2 Genes
Examkrackers MCAT Comprehensive Course, Charles Feng
[Link] fenguin@[Link] (224) 532-0039
Initiation
a. mRNA recognized by small subunit of ribosome, binds together
b. tRNA with 5-CAU-3 anticodon (bound to Met) binds to start codon at P
site of ribosome
c. Large subunit binds
Elongation
a. tRNA binds to A site of ribosome
b. Peptide bound to tRNA at P site gets attached to amino acid at A site
c. Translocation occurs; tRNA at P site shifted to E (exit) site and leaves,
tRNA at A site now attached to peptide moves to P site
Termination
a. Stop codon reaches A site
b. Release factors recognize it and cause disassembly of ribosome
Post-translational modifications can also occur where other stuff is attached
to amino acids
o If ribosome stays floating in cytosol protein ends up in cytosol
o If peptide contains a special signal peptide, a SRP will carry ribosome &
peptide to a receptor on the endoplasmic reticulum, causing the
ribosome to attach protein ends up inside ER, will become membrane
bound proteins or exported from the cell
Mutations
Gene mutation: change in DNA sequence within a gene
Chromosomal mutation: change in chrosome structure
Mutagen: something that increases the chance of mutation
Forward mutation: mutated organism mutating again
Backward mutation: mutation that cancels another mutation
Wild type: non mutated organism
Page 4 of 7
Biology Lecture 2 Genes
Examkrackers MCAT Comprehensive Course, Charles Feng
[Link] fenguin@[Link] (224) 532-0039
Gene Mutations
o Point mutation: one base pair changes, i.e. base-pair substitution, insertion/deletion
o Missense mutation: a base pair mutation that occurs in an exon, effects will vary
o Frameshift mutation: if insertion/deletion occurs in multiples other than 3 normally
results in nonfunctional protein
o Nonsense mutation: any mutation that creates a
stop codon nonfunctional protein
Chromosomal Mutations
o Deletions and duplications
o Translocation: segment from one chromosome
inserted into another
o Inversion: orientation of DNA reversed on a
chromosome
o Transposons are DNA segments that are able to
excise themselves from chromosome, reinsert
themselves at another location
Cancer
Unrestrained/uncontrolled growth of cells
Normal cell growth genes can be converted into oncogenes by carcinogens
Chromosomes
DNA wrapped up by histone proteins to form nucleosomes
Nucleosomes then coil up into chromatin
Humans have 46 separate dsDNA molecules, each called a chromosome
Each chromosome has a partner that codes for the same traits called a homologue
o Therefore humans have 23 homologous pairs of chromosomes
Diploid (2n): a cell that has homologous pairs - all somatic cells have 23x2=46
chromosomes
Haploid (n): a cell that doesnt have homologous pairs - all germ cells have 23
chromosomes
Cell Life Cycle
Cell life cycle: Go Sally, Go Make Children or G1 -> S -> G2 -> Mitosis -> Cytokinesis
G1: cell grows in size, can lead to G0 which is nondividing stage
S: DNA replicates, chromosomes change from looking like > to looking like X
G2: organelles duplicate, cell prepares to divide
M: cell divides
C: cytoplasm splits in two
Page 5 of 7
Biology Lecture 2 Genes
Examkrackers MCAT Comprehensive Course, Charles Feng
[Link] fenguin@[Link] (224) 532-0039
Mitosis - 1 diploid parent 2 diploid daughter cells
Prophase - chromatin condenses into chromosomes, centrioles
move to opposite sides, mitotic spindle begins to form,
microtubules from each centriole attach to opposite sides of
chromosomes at the kinetochore, nuclear membrane disappears,
nucleoli disappear
Metaphase - chromosomes align at middle of cell
Anaphase - chromosomes split in half
Telophase - chromatin decondenses, nuclear membrane reforms
Cytokinesis - cytoplasm separates into two cells
Meiosis - 1 diploid parent 4 haploid gametes/germ cells
After S phase: primary spermatocyte/primary oocyte
Meiosis I - primary spermatocyte turns into two haploid secondary spermatocytes, primary oocyte turns into one
haploid secondary oocyte, polar body
Meiosis II - 2 secondary spermatocytes turn into 4 spermatids. Only happens after fertilization for secondary oocyte,
which turns into a diploid zygote + another polar body
Prophase I: homologous chromosomes line up next to each other to form tetrads (XX), crossing over or genetic
recombination occurs
Metaphase I: XX lines up at equator
Anaphase I: two Xs split, travel towards opposite ends of cell
Telophase I: nuclear membrane may reform
Cytokinesis: primary spermatocyte splits evenly into secondary spermatocytes, but primary oocyte splits into a very
small polar body and large secondary oocyte
Prophase II/Metaphase II/Anaphase II/Telophase II occur similar to mitosis
Nondisjunction: if centromere of any chromosome doesnt split in Anaphase I or II, results in cells having not
enough/extra chromatids, common source of genetic diseases
Page 6 of 7
Biology Lecture 2 Genes
Examkrackers MCAT Comprehensive Course, Charles Feng
[Link] fenguin@[Link] (224) 532-0039
DNA Technology
Heating/adding salt can cause denaturation, higher % G-C needs high temp
Nucleic acid hybridization used to identify unknown sequences by binding
known sequences
Restriction enzymes
o Restriction enzymes cut DNA at specific palindromic sequences
o Can combine DNA cut with the same restriction enzymes to make
recombinant DNA
o Can add recombinant DNA to bacteria using a vector such as a plasmid or
a virus, replicate a bunch of times to produce a clone containing the DNA,
clones can be added to a clone library
o If recombinant DNA contains an antibiotic resistance gene, the resulting
clones can be screened by adding the antibiotic and keeping whats left
o Can also search a library for a desired DNA sequence by adding a probe, a
labeled complement to that sequence
Polymerase chain reaction: can amplify specific
double stranded DNA sequence using many copies
of DNA primers. Process multiplies the # of desired
dsDNA by two every time
Southern blot: identifies specific sequences of DNA
by chopping up DNA, separating pieces using gel
electrophoresis, then adding probe
Northern blot: same as Southern except for RNA
Western blot: detects specific proteins using antibodies
To replicate eukaryotic genes in bacteria, must start with eukaryotic mRNA which doesnt have introns, make
complementary DNA (cDNA) with reverse transcriptase then add it to the bacteria
RFLP analysis: identifies specific people based on distances between restriction sites
Page 7 of 7
