BIOTECHNOLOGY
Biotechnology uses living organisms to create products or facilitate processes
i.e. production of HGH verses extraction from cadavers
TOOLS
 Restriction Enzymes proteins made by bacteria that can cut DNA into pieces (bacterial defense)
Highly specific: cut nucleic acids at recognition sequences
(i.e. BamH1 cuts only at GGATCC) 1000’s exist
Sticky end: allows pasting two DNA fragments together
 Ligation is a process of permanently attaching two pieces of DNA by base pairing of sticky ends
 Plasmids : small circular pieces of DNA not a part of bacterial chromosome
Can replicate independently of the chromosome
 Transformation: uptake of new pieces of DNA into bacterial plasmids ( both plasmid DNA or recombinant
DNA)
 Cloning: To make an exact copy of a gene, cell, or organism. Allows
Creating organisms containing specific genes or livestock to serve as organ donors or blood
donors
 PCR: polymerase Chain Reaction is used to make large amounts of a specific piece of DNA from a very
small sample
 Recombinant DNA is DNA from 2 or more sources
Recombinant DNA Technology: Using the above tools, genes are combined from two or more different sources. The
recombinant fragment is introduced into a cell that can express that gene. Uses:
 Mass production of biochemicals needed by other species
 Creation of new strains of living organisms
 Production of specific protein sequences
Cloning a Protein
 Cut out gene for the protein of interest using restriction enzyme
 Insert that gene into a plasmid
 Transform the bacterial cell with that recombinant plasmid
 Allow the bacteria to reproduce itself and the plasmid.
 Harvest and purify the protein made in the bacterial cell
Cloning the Organism “Dolly”
1. An udder cell was isolated from a sheep and grown in culture (replicated)
2. An egg was taken from another sheep and its nucleus (DNA) was removed
3. The two cells were fused by electricity. Simulating a fertilization event only in this case the DNA is from one
parent.
4. The embryo was implanted into a surrogate mother sheep  Dolly (with the exact DNA from the original udder
cell.
Gel Electrophoresis: a technique that allows separating and sorting proteins and nucleic acid sequences based on
their size and charge.
Nucleic Acid sequences can be cut using restriction enzymes. Negatively charge fragments move to the
positive pole of the gel and separate according to size.
Longer macromolecules move through the gel more slowly than do shorter molecules.
No two individuals (except identical twins) has the same DNA sequence on homologous chromosomes.
Fragments (RFLP: restriction fragment length polymorphisms) differ in length and number of fragments produced ,
will migrate different distances in electrophoretic gel.
USES:
Forensics: DNA fingerprints uses RFLP (random fragment length polymorphisms) analysis. No two
RFLP’s are alike.
Paternity Cases
Evolutionary Relationships
Human Genome Project
Involves sequencing the entire genome: this provides the exact order of nucleotide pairs in each fragment and
chromosome, physical mapping each gene
Genome represents all genes present in a organism. Human genome is about 3 billion nucleotide pairs of DNA,
most does not code for genes
Human Genome is 1000 X larger that that in E. coli
Human G. has about 50000-100000 genes E. coli 2000 genes
97% is non-coding consisting of promoter and enhancers
Junk DNA include introns and non-coding regions between genes
Benefits: insight into embryo development; evolution, identification of genes that cause genetic disorders
and genes in common diseases like cancer
GMO= genetically modified foods