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Biotechnology has been defined as
the use of living things or parts of things to create or modify drugs and other substances, to modify food crops or other macroscopic organisms
Biotechnologies used for centuries include: (4)
- 1) fermentation
- 2) baking (saccharyomyces cerevisiae)
- 3) selective breeding
- 4) antibiotics
Nucleic acid therapeutics biotechnology includes:
- antisense nucleotides,
- DNA vaccines
Age of Modern Medical Biotechnology began:
- 1) 1982 � Human recombinant insulin (Humulin, E. Lilly) approved FDA
- 2) 1986 � Muromonab-CD3 (Orthoclone OKT3, Janssen), first monoclonal antibody approved by the FDA
emphasis of pharmaceutica biotechnology is on (3)
- 1) large molecule therapeutics
- 2) correcting gene mutation
- 3) replacement of damaged cells
�During 2009, the FDA and European Union regulators approved ___ new recombinant proteins (valued at ____).
The USDA regulates _______(3)
plants, plant pests, animal vaccines
The EPA regulates _____(4)
microbial/plant pesticides, microorganisms & animals that produce toxic compounds
The FDA regulates____(4)
food, human and animal drugs, human vaccines, transgenic animals
the lead regulatory agency in the United States is the
CBER is:____They regulate_____
�CBER: Center for Biologics Evaluation and Research regulates biotech products and other �biologics�
CDER is:_____ They regulate_____
�CDER: Center for Drug Evaluation and Research regulates traditional small molecule therapeutic agents.
FDA function include (3)
- 1) clinical development process
- 2) efficacy and safety
- 3) standards of manufacture, labeling, and distribution
Application to the FDA for biotherapeutics is called ___ _____ _____
Biologics License Application
biopharmaceutical is concerned with (2)
- 1) characterization of product
- 2) Contaminants not just restricted to small molecules
Transformation of Bacteria was first performe using 2 strains of ______
___ virulent strain that grew as _____ looking colonies on agar
_____ nonvirulent strain that produced ______ looking colonies on agar
in the Griffth Experiment, heated S cells mixed with R cells resulted in _____ mice
In the Griffth experiment, S cells contain a _____ _____ that can be transferred between cells
of the purified enzymes, only ______ treated smooth strains
Chargaff�s Nucleotide Complementarity states (2)
- 1.# of purines (A,G) = # of pyrimidines (C,T)
- 2.# of adenine = # of thymidine bases
_____ _____ _____allowed Watson & Crick to propose the double helix model that explained these identities.
Chargaff�s Nucleotide Complementarity
in order to manipulate DNA a series of _____ _____ enzymes are required
DNA manipulation enzymes (5)
- 1) DNA nucleases (DNase)
- 2) Restriction nucleases
- 3) DNA ligase
- 4) Reverse Transcriptase
- 5) DNA polymerase
_____ is any enyzme that is able to cleave the phosphodiester bond in DNA.
Deoxyribonuclease (DNase) comes in a Variety of specificities w/ some cleaving only _____ _____ DNA
DNA exonuclease � cleaves the _____ ___ ____ from the end of the DNA strand.
terminal (3') nucleotide
DNA ______ � cleaves within the DNA strand.
Mammalian _______ DNase has low specificity.
_____ ______are bacterial derived DNA endonucleases that possess unique specificities
It is believed that restriction nucleases evolved to protect bacteria from the insertion of _____ ___ into the chromosomal DNA
It was found that Restriction Nucleases could cleave DNA in a _____ number of _____ sequences.
Restriction nucleases recognize _____ in DNA seuences
Which of the following have overlap cutting? Arthrobacter luteus, Haemophilus aegyptius, Bacilus amyloliquefaciens H, Escherichia coli RY14, Haemophilus influenzae Rd?
The last 3
T/F the restriction enzyme is named from the source organism it cleaves.
true (ex. Alu for Arthrobacter luteus)
_____ is required in mammalian cells to ligate the Okazaki fragments during DNA replication
Selected ligases are able to work on _____ ended DNA fragments, but _____ fragments are more generally ligated.
Ligase forms a _______ bond, leading to a covalent bond, healing DNA cleavage points
____ ______ is the enzyme used to form a DNA strand using a template strand of DNA.
DNA Polymerase Always extends DNA strand in the _____ direction using a 3�-OH on a pre-existing primer piece of DNA.
5' to 3'
�DNA polymerase requires a _____
At least ___ types of eukaryotic DNA polymerase identified.
the primer strand of the polymerase reaction has a ____ for binding new bases
hydroxyl group at carbon 3
Prokaryotic DNA Polymerase 1 possesses three enzymatic activities:
- 1) 5' to 3' polymerase activity
- 2) 3' to 5' exonuclease activity (proofreading)
- 3) 5' to 3' exonuclease activity (DNA repair)
the Klenow fragment is cleaved from polymerase 1 by ______
Which enzymatic activity is the klenow fragment unable to perform that polymerase 1 can?
the 5� ? 3� exonuclease activity
In 1970, _____ _____ was isolated from RNA viruses that caused neoplasms in animals.
the RNA viruses that caused neoplasms in animals was ____ _____ _____
Rous sarcoma virus
Reverse Transcriptase is a ____ _____ DNA polymerase, meaning it uses _____ as a template for DNA synthesis.
RNA dependent, RNA
The enzyme from ____ _____ _____ _____widely used to convert RNA into DNA for manipulation
Moloney-murine leukemia virus (MMLV)
Using a series of highly purified enzymes, one can: (4)
- 1) Convert RNA into complementary DNA
- 2) Trim RNA to a particular size
- 3) Attach it to another DNA molecule
- 4) Make many copies of it in vitro