RECOMBINANT
DNA TECHNOLOGY
INTRODUCTION
RECOMBINANT DNA IS POSSIBLE
RECOMBINANT DNA : DNA DNA MOLECULES BECAUSE DNA MOLECULES FROM
FORMED BY LABORATORY METHODS
OF genetic
GENETIC recombination
RECOMBINATION (SUCH ALL ORGANISMS SHARE THE SAME
AS molecular
MOLECULAR cloning
CLONING) TO BRING CHEMICAL STRUCTURE. THEY
TOGETHER GENETIC MATERIAL FROM DIFFER ONLY IN
MULTIPLE SOURCES,
CREATING sequences SEQUENCES THAT WOULD NOT THE nucleotide
NUCLEOTIDE SEQUENCE WITHIN
OTHERWISE BE FOUND IN THE genome GENOME. THAT IDENTICAL OVERALL
STRUCTURE.
What is Recombinant DNA
• Recombinant DNA, which is often shortened to rDNA, is an
artificially made DNA strand that is formed by the
combination of two or more gene sequences.
•The technology used for producing artificial DNA
by combining different genetic materials (DNA)
Recombinant DNA Technology
from different sources is called Recombinant DNA
Technology. Recombinant DNA technology is
popularly known as genetic engineering.
•Using Recombinant DNA technology, we can
isolate and clone a single copy of a gene or a DNA
segment into an indefinite number of identical
copies.
•These new combinations of genetic material or
Recombinant DNA (rDNA) molecules are
introduced into the host cells, where they propagate
and multiply. The technique or methodology is
called Recombinant DNA technology.
Overview of Steps for developing rDNA.
• Isolation of genetic material from the organism.
• Cutting of vector DNA (plasmid DNA) and the gene of
interest (foreign gene) using the same restriction enzyme.
• Amplification of gene of interest using Polymerase Chain
Reaction (PCR).
• Adhering the foreign gene fragment with the cut vector using
the enzyme ligase. This leads to the formation of recombinant
vectors through a process called Ligation.
• Making the host cell competent and introducing the
recombinant vector into the host through a process called
transformation.
• Segregation of transformants and non-transformants through a
screening process, consequently obtaining the desirable gene
product.
Obtaining rDNA
Step 1: The DNA
fragment containing the
gene sequence to be Step 2: Cutting DNA.
cloned (also known as
insert) is isolated.
Step 4: Insertion of
these DNA fragments
Step 3: Joining DNA into host cell using a
“vector” (carries DNA
molecule).
Step 5: The rDNA
molecules are generated Step 6: Transfer of the
when the vector self rDNA molecules into
replicates in the host an appropriate host cell.
cell.
Step 8: Replication of
Step 7: Selection of the
the cells carrying rDNA
host cells carrying the
molecules to get a
rDNA molecule using a genetically identical
marker.
cell or clones.
Recombinant DNA cloning
Use of Recombinant DNA Technology
A plasmid made of
The DNA for insulin
Insulin Production DNA is removed
is first isolated
from a bacterial cell
A restriction enzyme
The insulin gene,
cuts the plasmid
with complementary
DNA open, leaving
sticky ends is added.
sticky ends.
Uses of Recombinant DNA Technology
DNA ligase enzyme The plasmid (now The bacterium host
splices (joins) together genetically modified) cell, divides and
the plasmid DNA and is inserted back into produces copies of the
the insulin DNA. the bacterium. plasmid.
The Bacterium makes The insulin is extracted
human insulin using from the bacterial
the gene in the
plasmid. culture.
Applications of Recombinant DNA
Technology
Preparation of gene maps.
In revealing details of various infections, diseases such as "inborn errors
of metabolism."
Finding out the complete nucleotide sequence of genome of an organism
and identification of genes.
Detecting cytogenetic abnormalities e.g. Down's syndrome, multifactorial
disorders, atherosclerosis, coronary artery disease etc.
Preventing various genetic disorders e.g. inherited haemoglobin
disorders, phenylketonuria, retinoblastoma etc.
Understand a molecular event is biological processes like growth,
differentiation, ageing etc.
Gene Therapy: Removal
Replacement or Production of and replacement of
correction of genetically defective genes with
normal healthy functional
deleterious modified organisms genes is known as gene
mutation by (GMOs) or therapy e.g. Sickle cell
transfer of clone transgenic anaemia, Severe
Combined Immuno-
gene in a patient. organisms for Deficiency (SCID).
providing particular
product and
nutrient.
• Applications in Medicine
Insulin production for diabetes treatment
Gene therapy for genetic disorders
Development of vaccines (e.g., Hepatitis B)
Applications in Agriculture
•Genetically modified crops (e.g., Bt cotton, Golden
rice)
•Pest resistance and improved yield
•Visual: Side-by-side comparison of regular vs GM
crops
Ethical and Safety Considerations
• Ethical concerns:
• Manipulating life forms
• Designer babies debate
• Safety concerns:
• Environmental impact
• Potential for misuse
Future Prospects
•Personalized medicine
•Advances in synthetic biology
•Solving food scarcity
Conclusion
•A revolutionary tool for science
•Benefits outweigh challenges
•Promises a better future for health,
agriculture, and the environment
