Chapter 1 notes

groupmoujeanteaBiotechnology

Oct 23, 2013 (3 years and 9 months ago)

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Chapter 1

Biotechnology and


the Workplace

Introduction


Biotechnology


The use of organisms/materials derived from organisms


Make useful products


Not a recent phenomenon


Used organisms to make products


Wine, cheese, bread


Usually referring to something mor
e “dazzling


Cloned sheep, gene therapies, DNA fingerprints


Introduction


Modern biotechnology


Rooted in basic laboratory research


Broadens/deepens understanding of how living organisms work


The application of knowledge useful to humans


Knowledge gained app
lied to create modern biotechnology


Technologies of Modern Biotechnology


Modern biotechnology


Manipulation of DNA


Tools cut DNA at specific sites


Enzymes to splice DNA strands together


Techniques to visualize DNA


Techniques to separate DNA fragments from
one another


Techniques to identify fragments of DNA with SPECIFIC sequences


Techniques to amplify DNA


Techniques to sequence and synthesize DNA


Technologies of Modern Biotechnology


Tools and the terms


Allow scientists to introduce genetic information/DNA
from one organism to another


Genetically modified/genetically engineered


When a biologist causes a cell or organism to take up a gene from another organism










Technologies of Modern Biotechnology


More terms


Recombinant DNA


DNA that contains sequence
s of DNA from different sources that were brought together
using the tools of biotechnology


Often referred to as “revolutionary”


There is POWER in the manipulation of DNA!


DNA directs the cell in making a specific protein


Called expression (the protein is
expressed)


Technologies of Modern Biotechnology


How is DNA transferred?


Plasmids


Plasmids are molecules of DNA that are found in bacteria separate from the bacterial
chromosome. They:


are small (a few thousand base pairs)


usually carry only one or a few

genes


are circular


have a single
origin of replication


Act as a vector


Carries a desired gene into the recipient cell


Other vectors include viral vectors and direct injection


Under proper conditions, plasmids are readily taken up by recipient bacterial
cells


Technologies of Modern Biotechnology


After bacteria transformed, then what?


Fermentation


The large
-
scale cultivation of microorganisms


Alter the bacteria


Grow in media


Harvest the protein of interest


What about other types of cells?


Possible to intr
oduce gene of interest into cultured mammalian cells/other multicellular
organisms


Cultured cells


Cells grown in flasks, dishes, vats, or other containers outside a living organism


Grow more slowly than bacteria


More fragile


More complex nutrient requireme
nts


Introducing gene of interest into plants and animals


Transgenic


A plant or animal whose cells are genetically modified using the techniques of
biotechnology




Technologies of Modern Biotechnology


Genes responsible for particular traits or disease sus
ceptibility are chosen and extracted.


Next they are injected into fertilized mouse eggs.


Embryos are implanted in the uterus of a surrogate mother.



The selected genes will be expressed by some of the offspring


Technologies of Modern Biotechnology


What

about humans?


Raises critically important societal and ethical issues


Can it be passed to offspring


Present research into curing illness


Intended to cure individuals


Not affect children of the treated person, just the illness or condition


What use then?


E
xplore role of genes


Mechanisms of control


Technologies of Modern Biotechnology


Other uses proving to be beneficial to humans


Monoclonal antibodies


Antibodies


Proteins of the immune system


Recognize/bind to substances invading the body


Bind only to a spec
ific target


Exceptionally homogeneous populations of antibodies directed against a specific target


Produced by hydridoma cells


Result from the fusion of an antibody
-
production cell and a cultured cell


Very specific and useful


Example: home pregnancy tests


Applications
-
Medical/Veterinary


Biopharmaceuticals


Pharmaceutical products, like insulin or Factor VIII, that are manufactured using
genetically modified organisms as production systems


Transgenic animals


Can produce desired protein


Typically secreted in
to milk


Isolated and purified


Transgenic plants


Genes introduced into crop plants


Cultivated and harvested to obtain product of interest


Gene therapy


Involves replacing a gene that is missing, or correcting the function of a faulty gene, in
order to treat
or cure an illness



Examples
-
E. coli


Recombinant DNA products produced in
E. coli


Human insulin
-
1982


Before purified from pancreas glands of animals for human consumption


Genetically modified


More reliable source


Less likely to cause allergic reactions


Hu
man Growth Hormone
-
1985


Before, dwarfism treated with hormone purified from pituitary glands from cadavers


Some using this died from neurodegenerative disease
-
Creutzfeldt
-
Jakob disease!!!!!!


Interferon
α
-
2b
-
1986


Treats variety of viral diseases and cancers


Hairy cell leukemia, AIDS
-
related Kaposi’s sarcoma, renal cell carcinoma, and chronic
hepatitis B


Known about in 1957, but difficult to isolate (viral disease in patients, ONLY)


Not available until recombinant methods developed


Examples
-

Yeast Cells


Reco
mbinant DNA products produced in yeast cells


Hepatitis B vaccine
-
1986


Viral illness
-
no treatment currently available


Before recombinant methods, vaccine prepared from plasma of hepatitis
-
infected
humans


Limited sources


Concerns about purity


From yeast cell
s


Unlimited sources


Reduced possibility of contamination


Examples
-

Mammalian cells in culture


Recombinant DNA products produced in mammalian cells in culture


Erythropoietin
-
1989


Used to treat anemia from renal failure or AZT treatment


EPO produced in the
kidney


Stimulates production/maturation of rbc’s


Found in small quantities in urine


Never available before recombinant methods


Anemia from kidney disease treated with transfusions



Factor VIII


Treatment for hemophilia


Before recombinant methods, source was

human plasma


Patients contracted AIDS




Applications
-

Agricultural/Food Related


Breeding programs for plants and animals


Enhance characteristics


Benefit from biotechnological methods


Very specific genetic manipulation organisms


Does away with most undesi
rable traits passed on by traditional manipulations


VERY controversial


Not as safe as conventional foods?


Introduction of genetically modified organisms into environment may have unforeseen
and adverse effects


Benefits


End severe food shortages


Enhance nut
rition of plants and animals


Reduce reliance on chemical pesticides


Other Applications


Bioremediation


Clean
-
up of contaminated soil and water


Identification of individuals


Human DNA sequences very distinct (except for identical twins)


DNA fingerprinting


The Organization of a Biotechnology Company


Research and Development
-

R&D


Find a potential product WITH commercial value


Performs research relating to the potential product


Characterizes properties of the product


Composition, physical and chemical properti
es


Strength, potency, or effect of the product


Purity of product


Steps required to avoid contamination


Stability/shelf
-
life of product


Applications of product


Safety concerns in use of the product


Establishes product specification


Properties that every bat
ch of final product must have to be released for sale


Develop testing methods to ensure meets specifications


Develop processes to make product


Describe cells or microorganisms to make product


Raw materials needed and specifications of those materials


Equip
ment needed


Plan for production



The Organization of a Biotechnology Company


Production Unit


Make product


Scale
-
up


Work with large
-
scale equipment/large volume reactions


Monitor and control environment as required for the product


Proper temperature, steri
lity, etc.


Cleaning, calibration, and maintenance of equipment


Follow written procedures associated with producing product


Monitor processes


Initiate corrective actions if problems arise


Complete paperwork!!!!!! (ALWAYS lots’o this)


The Organization of a
Biotechnology Company


QA/QC


Monitor equipment, facilities, environment, personnel, and product


Reviewing all production procedures used in the company


Ensure accuracy of all documents


Test samples of product and materials need for producing product


Compare

data to established standards


Decide whether or not to approve product for release


Review customer complaints


The Organization of a Biotechnology Company


Other functions


Engineering or facilities management


Ensures systems that control building are oper
ating properly


Large equipment properly installed and functioning


Facility maintenance and housekeeping


Receiving and shipping


Dispensing


Metrology


Marketing and sales














Different Types of Biotechnology Work Environments


Basic Biological Researc
h Labs in Academic or Government


Basic research


Investigate fundamental problems in biology


Product is knowledge or information


May result in a commercial product


R&D laboratories associated with industry


Investigate questions in biology intended to result

in commercial products


Production facilities


Make products


Usually use large scale equipment


Testing laboratories


Analytical, quality control, forensic, microbiology, metrology, clinical testing labs


Different Types of Biotechnology Work Environments


Reg
ardless of work environment


Scientists and technicians involved in the physical aspects of acquiring data or making a
product


Grow bacterial cells


Plants


Animals


Work with materials derived from organisms


Make measurements, prepare reagents, gather/record data