Applications of

Recombinant Technology
A/P Yeong Foong May
Dr Lee Zheng Wei
Topics

• In this lecture, we will discuss more in depth about

recombinant technology
• We will explore examples of application of recombinant
technology in various fields
• We will discuss what the scientists did as a community
at the beginning when they realized they can manipulate
DNA
Learning outcomes

• At the end of this week, you should be able to

• Describe the applications of recombinant technologies
• Discuss scientists’ contributions to regulating the use of
recombinant technologies in the early days
Applications
• From the fundamental understanding of DNA, scientists found ways to
apply their knowledge to improving different areas of our lives.
• The field of recombinant technology has contributed to:
• Industrial Biotechnology
• Pharmaceutical and Medicinal Products
• Commercial Agriculture
 Recap: A typical workflow of recombinant DNA technology
Plasmid
1 Vector
PCR amplified fragment
1. Obtain and amplify DNA fragment encoded for the
2
Restriction
desired gene product. Enzymes

2. ‘Cut’ and ‘paste’ the DNA fragment into a plasmid Digested

Digested fragment Vector
vector.
3. Introduce the recombinant plasmid DNA into the host 2
Ligation Recombinant
cell. Plasmid
DNA
4. The host cell would be able to produce the desired Ligase
3
gene product with its own cellular machinery. 4
Transformation
Why recombinant technology?
• Specific and speedy amplification of gene fragment using polymerase chain reaction (PCR)
• Precise cutting and pasting of gene fragment using restriction enzymes
• (Unlimited) possibility to create different recombinant DNA encoding for desired gene product
• Exemplar of applications of such technology in various fields:
A. Industrial Biotechnology
B. Pharmaceutical and Medicinal Products
C. Commercial Crops in Agriculture
A. Industrial Biotechnology
• Food processing
• Genetically engineered microbial to produce enzymes used to process foods
• E.g. lactase to produce lactose-free milk
• Biofuel production
• Bioethanol produced from fermentation of sugar or starch by microbes
• Environment maintenance
• Microbial degradation of hydrocarbons in oil spillage and plastics

Alcanivorax borkumensis
B. Pharmaceutical and Medicinal Products
• Vaccine development and production
• Molecular cloning of targets and laboratory testing
• Synthetic biology
• Natural active compounds originated from algae, fungus and plants
• Novel directed or random mutagenesis of protein enzymes
• Cell-free system production
• Assemblies of proteins required for compound production in test
tube
C. Commercial Agriculture
• Over 100-fold increase in production of Genetically Modified (GM) crops
in the past two decades.
• Soybean (~50%), maize (~30%), cotton (~13%) and canola (~5%) are 4 major GM
crops.
• United Nation (UN) Cartagena Protocol (2000):
• “living modified organism” = novel combination of genetic material obtained
through the use of modern biotechnology (in vitro nucleic acid techniques
including recombinant DNA, or fusion of cells beyond taxonomic family)
https://www.cbd.int/doc/legal/cartagena-protocol-en.pdf
 Plant Breeding Methods
Selective Breeding Mutation Breeding Transgenic Breeding
Plasmid

Targeted foreign gene

Chemical or radiation insertion
Select and backcross Select and backcross Select and culture
over many generations ~6-7 years ~4-6 years
~8-10 years or more
Implications of
Recombinant Technology
Knowing what we know now about the powers of recombinant DNA
technology, it is important to know that at the beginning when scientists first
discovered that they were able to manipulate DNA that there were not only
different views as to the extent they should go with the technology, but that
there were efforts by the scientists who pioneered the field to impose self-
regulation on themselves in terms of the types of experiments they allowed
themselves to carry out

1970 Paul Berg (Nobel Laureate;
‘father’ of recombinant DNA
technology) and David Jackson
ligated together DNA from SV40
(simian virus) and lambda phage
(bacterial virus)
Berg self-imposed a
moratorium on experiments
involving DNA hybrids -
these hybrids will not be Asilomar Conference with a wider
propagated in cells but will Pre-Asilomar meeting among key scientific community and
remain only in test-tubes scientists outsiders

1970 1971 1973 1975

Brief sequence of
events
1971 1973 1974

Janet Metz, a graduate student of
Paul Berg presented her idea to
cut and paste viral and bacterial
DNA together*
Herbert Boyer and Stanley Cohen
cut and pasted together DNA
fragments from two different
bacteria with one fragment
containing antibiotic resistance
‘Berg Letter' published on the
restrictions or moratorium on the
use of certain kinds of
recombinant technology

* This triggered other scientists at the presentation to raise the alarm bells on the dangers of genetic hybrids
Essential Videos and Reading
Paul Berg (Stanford) - Recombinant DNA and Science Policy:
https://youtu.be/QSKe15I4vyM

Berg et al, 1974, Potential hazards of recombinant DNA molecules, Science , Jul. 26, 1974,
New Series, Vol. 185, No. 4148 (Jul. 26, 1974), p. 303.
Berg et al, 1975. Summary statement of the Asilomar conference on recombinant DNA
molecules. Proc. Nat. Acad. Sci. USA Vol. 72, No. 6, pp. 1981-1984, June 1975.
- I have uploaded these at Canvas