BIOTECHNOLOGY REVIEWER

1. Chromosome, DNA, Gene

Chromosome a thread-like structure of nucleic acids and protein
found in the nucleus of most living cells, carrying genetic
information in the form of DNA
DNA deoxyribonucleic acid; self-replicating material which is
present in nearly all living organisms as the main constituent of
chromosomes, it is the carrier of genetic information
Gene unit of heredity which is transferred from a parent to
offspring
-DNA- in early 1950s
a. James Watson
b. Francis Crick
3 complex sub-units of nucleotides in DNA
Phosphate
Pentose or Deoxyribose
Nitrogenous Bases

2. Central Dogma of Protein Synthesis

Helicase the enzyme that uncoils the strands
*semi-conservative strands*
Replication (DNADNA) Transcription (DNAmRNA) Translation (mRNAtRNA)

Hydrogen Bond Bonds the bases

G+C triple hydrogen bond
A+T double hydrogen bond
*DNA will exit the nucleus and enter the cytoplasm where the message is
passed to tRNA
LAGGING STRAND and LEADING STRAND
Okazaki Fragments (the spaces between) and the RNA Primers
Primase, Ligase and RNA Polymerase are enzymes that try to connect

3. Polymerase Chain Reaction

Process of amplifying DNA exponentially
Can be applied to plants, virus, bacteria, etc.
Related Terms
Template DNA contains the sequence of DNA you want to
amplify
DNA Polymerase otherwise known as Taq (Thermus
Aquaticus) Polymerase, it is an enzyme that makes a new
strand of DNA
Primers are small segments of single stranded DNA which
binds to a specific region on either side of the target DNA (20
nucleotides long)
Buffer is a salt solution that helps to stabilize the DNA and
other components of the reaction (an enhancer)
Steps
a. Denature DNA DNA is heated to 95C. This breaks the hydrogen
bonds that hold DNA strands in a helix, allowing the strands to separate
creating single-stranded DNA
b. Primer Annealing the mixture is cooled to anywhere from 45C to
72C. This allows the primers to bind or anneal to their complimentary
sequence in the template DNA

c. Extension the reaction (DNA) is then heated to 72C, the optimal

temperature for DNA Polymerase to act. DNA polymerase extends the
primers, adding nucleotides onto the primer in a sequential manner,
using the target DNA as a template

4. Genomic Library
is a collection of bacteria which have been genetically engineered to
hold the entire DNA of an organism
is a collection of genes or DNA sequences created using molecular
cloning
constructed using clones of bacteria or yeast that contain vectors into
which fragments of partially digested DNA have been inserted. These
bacteria and yeast are subsequently grown in culture and when these
microorganisms replicate their genome, they also replicate the vector
genome contained within them (that is, they replicate DNA fragments
that had been inserted in vectors producing clones of the original
genome)
Steps involved in the construction of genomic library
a. To create a human genome library, a researcher begins by
extracting and purifying DNA from human cell
b. The purified DA consists of extremely long strands, the strands are
cut into manageable sizes
c. DNA is digested with restriction enzymes which cut the DNA into
thousands of smaller fragments at specific sequences, each of which
contain one or more gene
d. Each fragment must be inserted into loops of DNA called plasmids
e. Plasmids are digested with the restriction enzymes and then sealed
to human DNA using DNA ligase enzyme, the resulting molecules are
recombinant
f. Recombinant DNA molecules are added to bacteria, and the bacteria
are made to take up the DNA. When bacteria have taken up the DNA,
the entire collection of cells and DNA represents a human genome
library.

Probe is normally a cloned piece of DNA that contains a portion of

the sequence for which you are searching

5. Other Terms and Definitions

ETHIDIUM BROMIDE - a molecule that binds to DNA by intercalating
(?) between the bases and fluoresces in the presence of ultraviolet
light, used as a stain for DNA, especially agrose gels
FNRI - Food and Nutrition Research Institute
INSULIN - A hormone secreted by the B (Beta) cells in the pancreas
that reduces glucose level in the body

CDNA - Collection of expressed mRNA's usually from specific cells in

an organism
INTRON - a non-coding gene that is removed during RNA processing
after transcription
BOTOX - a trade name, derived from Botulinum toxin, a protein and
neurotoxin produced by the bacterium clostridium botulinum
HIV - Human Immunodeficiency Virus
IRRI - International Rice Research Institute
GEL ELECTROPHORESIS - A method used to separate DNA or RNA
fragments by length protein can be separated by size/charge
FREDERICK SANGER - Proponent of a DNA Sequencing technique in
early 1970s