CELL TRANSPORT

A. Passive
1. Passive Diffusion – higher to lower concentration
2. Osmosis
a. Isotonic solution – water moves in and out of the cell
b. Hypertonic solution – water moves out of the cell
c. Hypotonic solution – water moves into the cell
3. Facilitated Diffusion
B. Active – requires use of ATP; energy driven process
Two Types:
a.) Primary Cellular Transport – transport sodium and potassium
b.) Secondary Cellular Transport – transport amino acid and glucose
1. Pump – specific for sodium potassium ions pump; from low to high concentration
2. Vesicle Transport

Protein Synthesis
A. Transcription = nucleus
- First part of the central dogma of molecular biology: DNA -> RNA (mRNA)
B. Translation = cytoplasm
- Second part of the central dogma of molecular biology: RNA -> protein
Transcription Flowchart
- DNA double helix in the nucleus -> DNA unwinds -> Complementary strand of mRNA made ->
Complementary strand of mRNA processed -> mRNA exported into cytoplasm and DNA helix reformed
Codon Table

AMINO ACID FUNCTIONS

phe Phenylalanine - Converted into the
neurotransmitters
dopamine,
norepinephrine, and
epinephrine
- Serves as precursor for
other important
molecules like tyrosine
leu Leucine - Muscle building
- Helps regulate blood
sugar levels and provides
energy for the body
ile Isoleucine - Energy production
- Regulation of blood sugar
level
met Methionine - Precursor for cysteine,
carnitine, and creatine
- Plays a role in
detoxification
val Valine - Helps maintain proper
nitrogen balance in the
body
pro Proline - Structure and stability of
protein
- Important component of
collagen
thr Threonine - For production of
antibodies and enzymes
- For metabolism of fats
 ala Alanine - Synthesis and
metabolism of amino
acids and glucose
tyr Tyrosine - Involved in production of
thyroid hormones and
the pigment melanin
his Histidine - Involve in maintaining pH
balance in the body
gln Glutamine - Maintaining integrity of
intestinal lining
asn Asparagine - Metabolism of ammonia
- Formation of
glycoproteins
lys Lysine - Collagen formation,
hormone production, and
immune function
- Calcium absorption
asp Aspartic Acid - Energy metabolism
glu Glutamic Acid - Important
neurotransmitter in the
brain
cys Cysteine - Production of
glutathione
trp Tryptophan - Production of niacin (Vit.
B3)
arg Arginine - Wound healing
- Precursor for nitric oxide
gly Glycine - Supports the production
of other important
molecules like heme

Splicing – removal of non-coding sequences

5’ Capping – ensures mRNA’s stability
Polyadenylation- helps mobility of mRNA

Mature RNA – carries genetic message from nucleus to cytoplasm in the form of codons

Translation Flowchart
- mRNA binds to ribosomes in cytoplasm / RER -> 2 tRNA anticodons H-bond with mRNA codons -> Peptide
bonds form between adjacent amino acids -> Ribosome moves along mRNA strand to next triplet codon ->
tRNA released -> Polypeptide chain produced -> Polypeptide folds into PROTEIN
Initiation – transitional complex forms, and tRNA brings first amino acid in polypeptide chain
Elongation – tRNAs bring amino acids one by one to add to polypeptide chain
Termination – release factor recognizes stop codon

CELL REPRODUCTION
Cell Division – process by which a cell divides to form two new cells (daughter cells)
 Three types of cell division / reproduction
o Prokaryotes (bacteria)
 Binary Fission – divides forming two new identical cells
o Eukaryotes
 Mitosis
 Cell or organism growth
 Replacement or repair of damaged cells
 Meiosis
 Formation of sex cells, or gametes
WHY DO CELLS DIVIDE?
1. DNA Overload
- If cells grow without limit, “information crisis” would develop
- DNA cannot serve the increasing size of cell
2. Exchange of materials
- Food and oxygen have to cross membrane very quickly
- Waste must get out
- If cell is too large, this occurs slowly and the cell will die

PROKARYOTIC CELL DIVISION

 Binary Fission
 o 3 main steps:
1. DNA Replication – DNA is copied, resulting in 2 identical chromosomes
2. Chromosome Segregation – 2 chromosomes separate, moves towards end poles of cell
3. Cytokinesis – cytoplasm divides, forming 2 cells
 Each daughter cell is genetically identical to parent cell

EUKARYOTIC CELL CYCLE

 Interphase – period of growth and DNA replication between cell divisions
o Three phases:
 G1 Phase – cell increases in size
 S Phase – replication of hormones; two strands called sister chromatids joined by centromere
 G2 Phase – organelles double; new cytoplasm forms; all other structures needed for mitosis form

TWO MAIN STEPS OF CELL DIVISION

1. Mitosis (4 steps: Prophase, Metaphase, Anaphase, Telophase)
2. Cytokinesis – cytoplasm divides, forming 2 cells
 Each daughter cell is genetically identical to parent cell

MITOSIS
 Process that divides cell nucleus to produce two new nuclei
 Continuous process – occurs in somatic cells
 Four phases (PMAT)
o Prophase
1. Chromosomes visible
2. Centrioles migrate to the pole
3. Nuclear membrane disappears
4. Spindle forms
o Metaphase
1. Chromosomes line up on the equator of the cell
2. Spindles attach to centromeres
o Anaphase
1. Sister chromatids separate
2. Centromeres divide
3. Sister chromatids move to opposite poles
o Telophase
1. Chromosomes uncoil (now chromatin)
2. Nuclear membrane reforms
3. Spindle disappears

MEIOSIS
 Occurs in sex organs only
 Produces sex cells – cells with ½ the number of chromosomes as the original cell
 Males – meiosis produces 4 sperms; other 3 cells are called polar bodies – they give up their cytoplasm to
nourish the egg
 Females – meiosis produces 1 viable egg
 Phases:
o Prophase I – Crossing Over
1. Longest and most complex phase (90%)
2. Chromosomes condense
3. Synapsis – when homologous chromosomes pair up to form tetrads
4. Crossing over may occur in the tetrad
o Metaphase I
1. Shortest phase; paired homologous align
2. Independent Assortment – pairs of homologues line up independently of other pairs’ orientation
o Anaphase I
1. Homologous chromosomes separate towards the poles
2. Sister chromatids remain attached
o Telophase I
1. Each pole now has haploid set of chromosomes
2. Cytokinesis – two haploid daughter cells formed

DNA REPLICATION
DNA REPLICATION MITOSIS
PURPOSE Make an identical copy of DNA before Growth, repairs, and maintenance of
cell division occurs multicellular organisms
PROCESS Occurs during S phase Occurs during M phase
RESULT Two identical DNA molecules, each Division of two genetic material into
containing one. Original strand and two sets, ensuring that each daughter
 one newly synthesized strand cell receives a complete set of
chromosomes

DNA REPLICATION PROCESS

1. DNA double helix unwinds – initiated by helicase (breaks hydrogen bond)
2. Separation of two single strands of DNA creating a replication fork – strands will serve as a template for making a
new strand of DNA
a. Leading strand – 3’ to 5’ direction (towards replication fork)
b. Lagging strand – 5’ to 3’ direction (away from replication fork)

DNA PROFILING TECHNIQUES

1. Restriction Fragment Length Polymorphism (RFLP)
- DNA – restriction enzymes (endonucleases) = minisattelites/VNTRs (specific sequence)
- 4-6 polymorphic loci are analyzed – core sequence – 10-100
- 1980 – D14S1 – first DNA polymorphism suitable for forensic purposes
- Multicolor VNTRs – other hypervariable regions at several loci
2. STR Analysis (Short Tandem Repeats)
- Short sequences throughout human genome
How it works:
1. STR loci – targeted and amplified (PCR)
2. DNA fragments – separated and detected using electrophoresis
a. Capillary electrophoresis
b. Gel electrophoresis
3. Electropherograph – interpreted through statistical analysis

The more STR regions that are tested in an individual, the more discriminating the test becomes

3. Y-STR Analysis / Y Chromosome Analysis

- Y chromosome – str – males
Applications:
1. A mixture containing more than one male contributor
2. Sexual assault cases – evidence samples are more difficult to resolve with traditional autosomal STR
analysis
o Vaginal swabs containing mixed male and female DNA sample
3. Paternity testing
4. Identification of missing persons
5. Trace lineage if genealogical research
Disadvantages:
1. Useful for excluding a suspect but not powerful enough to identify a unique match
2. All males of a family will have exactly the same Y-STR profile
3. Y-STR typing results cannot be uploaded into the CODIS at this time

DNA EXTRACTION
- Removal of DNA from cells
- DNA isolation and DNA purification
Instrumentation use in DNA Extraction (BCG)
1. Bead Beater – “lysing” of cells
2. Centrifuge – to precipitate DNA
3. Gel Box – separate DNA in an agarose gel with electrical charge
3 Parts of DNA Extraction
1. Lysis (LPP) – breaking cells to release DNA
a. Cell wall / Membrane Lysis
i. Physical method – mechanical disruption
ii. Chemical method
Detergent – sodium dodecyl sulfate (SDS) / Lauryl Sulfate
Enzymes – amino acid bonds
Animal cells – protenaise K
Plant cells – cellulase
b. Precipitation – separates DNA from cellular debris

DNA EXTRACTION METHODS

1. Organic Method – yields highest quantity as quality
2. Chelex Method – used when samples contain very few cells
3. FTA Paper Method – used to collect reference sample
DO’S AND DON’TS IN THE HANDLING AND PRESERVATION OF BIOLOGICAL EVIDENCE FOR
DNA ANALYSIS

DO’S
1. Ensure proper chain of custody.
2. Properly preserve and document evidence for identification during court presentation.
3. Have separate containers for different samples.
4. Place blood or liquid tissues in spill-proof containers.
5. Immediately extract DNA samples from the biological evidence.
6. Materials in contact with the samples should be sterile.
7. Always wear gloves and additional protective device when appropriate.
8. Blood samples in liquid form must be stored frozen or placed in an ice pack.

DON’T’S
a. DNA test is very sensitive and vulnerable to contamination by other DNA sources. Avoid
contamination with other human DNA.
b. Do not handle samples with bare hands.
c. Avoid long-term storage of tissues.
d. No ice is required and dry ice should never be used to cool the tube of blood.
e. Do not mouth pipette anybody fluid regardless of the source.
f. Avoid drying of wet stained garments under direct sunlight or exposing it to an electric fan.
g. Do not collect a wet garment or garment with a wet bloodstain in a sealed air-tight container.
h. Do not use plastic bag in the collection of blood specimen.

PROTEINS – the basic substance of every body cell including the muscles, blood, skin, nails, hair, hormones and internal
organs
Amino Acids – the building blocks of proteins
a) Essential Amino Acids – 8 amino acids that cannot be synthesized by the body so they must be sourced from the food
that people eat
e.g. Isoleucine, Leucine, Lysine, Methionine, Tryptophan, Valine, Phenylalanine, Tyrosine
Non-essential Amino Acids - amino acids produced / synthesized by the body e.g. Serine, Proline, Cystine, Glycine,
Glutamic Acid, Histidine, etc.
Exons - stretches of DNA sequences that are represented in the mature form of RNA, including mRNA and tRNA
Introns - intervening DNA sequences between exons that will be spliced from the maturing RNA molecule
RNA Transcript - usually consists of multiple exons spliced together
Protein Synthesis - the process of making proteins
DNA Replication - the process by which identical copies of DNA are produced
Codons - 3 consecutive RNA nucleotides
Anti-Codons - 3 consecutive nucleotides that bond with the codon to form amino acids
Primers - short pieces of single-stranded DNA that are complementary to the target sequence
needed to start DNA replication / amplification process
DNA Helicase - the enzyme responsible for the unwinding of the double-helix DNA strand during replication process and
the production of a replication fork
RNA Polymerase - the enzyme responsible for splitting the double helix of DNA
DNA Primase - enzyme that produce a short piece of RNA strand called primer
Primer - acts as the starting point for DNA synthesis
Exonuclease - the enzyme that remove the primers
DNA Ligase - the enzyme that seals up the sequence of DNA into two continuous double strands
mRNA (messenger RNA) - carries genetic message from the nucleus to the cytoplasm in the form of codons for protein
synthesis
rRNA (Ribosomal RNA) - RNA found in the ribosome that allows interaction of mRNA and tRNA
tRNA (Transfer RNA) - bears the anticodon (complementary to the codon) to specify the amino
acid to be produced

Only 4 types of bases are associated with DNA:

a) Adenine (A)
b) Guanine (G)
c) Cytosine (C)
d) Thymine (T) – only present in DNA

Uracil – found in RNA in place of thymine

A and G = purines
C and T = pyrimidines
***Purines and pyrimidines are bases because their nitrogen atoms can accept protons.
***The human genome contains approximately 3 billion base pairs and their sequence regulates the expression of
genes.