DNA recombinant technology involves manipulating genes in organisms. It uses restriction enzymes to cut DNA at specific sites, vectors to carry foreign DNA, and host organisms for transformation. The key steps are: 1) Isolating the gene of interest from a library or by synthesis 2) Inserting the gene into a vector like a plasmid using enzymes and ligation 3) Transforming the vector into a host organism through physical or chemical methods 4) Selecting transformed host cells that contain the recombinant DNA 5) Analyzing the new genes and proteins produced by the host.

1. DNA Recombinant Technology
Advance Metabolic Engginering
(Course Code 157609
Sahri Yanti
Student ID: 10832910
Ph.D Program
Applied of Chemistry Departement
Chaoyang University of Technology, Taiwan

2. Definition
01
Tools of Recombinan
DNA
02
Step in Recombinan
DNA
03
Journal Example
04
OUTLINE

3. DefinitionRecombinan DNA
Tecnology
Genetic engineering
Experimental techniques to manipulation of
The genes in an organism
Molecular cloning and transformation
Involves the scientific alteration of the structure of genetic
material in a living organism
Order to produce new species or bio-chemicals
The development and application of scientific methods, procedures, and
technologies that permit direct manipulation of genetic
Scarce biological product by introducing DNA isolated from
animals or plants into bacteria and then harvesting the product
from a bacterial colony
Identifiation, isolation and Insertion of gene interest into a vector

4. GENE, DNA & RNA
DNA, RNA, Diferent funtion and structur of DNA vs RNA

5. ■ Gene and DNA are the two genetic terminologies mainly
used to determine the characteristics of a living thing
■ In general, a gene is a short section of DNA and DNA-
Deoxyribonucleic acid is a molecule which carries the genetic
instructions or the hereditary materials
DNA
Genes
Chromosomes
Cell & Nucleus
Gene DNA
Genes are the DNA
stretches
which are encoded for
different proteins
DNA is a chemical which
stores. the genetic
information of an
organism
Regulates the traits of an
organism
Regulates the gene
regulation.
A small stretches of
DNA.
A long chain
polynucleotide.
Genes are made up of
either DNA
or RNA.
DNA is made up of
simpler monomer units
called nucleotides
Are coded with heredity
information.
Encodes the genetic
instructions.

6.  There is another type of genetic material found in cells and
viruses known as ribonucleic acid (RNA).
 The genetic information stored in DNA is then transcribed
into RNA, and the details in RNA are then translated for the
synthesis of the protein (nucleoprotein)
DNA & RNA fungtion
DNA RNA
 Replication process:
Transferring the
genetic information
from one cell to its
daughters
 and from one
generation to the next
 Equal distribution of
DNA during the cell
division
 Mutations: The
changes which occur
during the DNA
sequences
 Transcription:
 Cellular Metabolism
 DNA Fingerprinting
 Gene Therapy
 Facilitate the
translation of DNA
into proteins
 Functions as an
adapter molecule in
protein synthesis
 Serves as a
messenger between
the DNA and the
ribosomes
 They are the carrier
of genetic
information in all
living cells
 Promotes the
ribosomes to choose
the right amino acid
which is required in
building up of new
proteins in the body.

7. DNA RNA
Deoxyriobosa as a suger Ribosa as a sugar
4 nitrogen bases in DNA:
Adenine, Guanine,
Cytosine, and Thymine
that pair together)
A—T ; C— G
4 nitrogen bases in RNA, A,G,C,
and U that pair together)
A—U ;C—G
DNA is DOUBLE
STRANDED, cannot
leave the nucleus. DNA
is tightly wound into
chromosomes. it is
exactly the same in each
cell. Portions of DNA are
called genes.
RNA is SINGLE STRANDED and
does not have to stay in the
nucleus. RNA not found in
chromosomes because it does
not carry the genetic code,
however it can read the DNA
code and take the information out
of the nucleus
DNA & RNA structure

8. The different types of RNA are involved in a various cellular process
tRNA
The transfer RNA is
responsible for choosing
the correct protein or the
amino acids required by
the body in-turn helping
the ribosomes.
rRNA
The rRNA is the
component of the
ribosome and are
located within the in the
cytoplasm of a cell,
where ribosomes are
found.
The ribosomal RNA is
primarily involved in the
synthesis and
translation of mRNA
into proteins in all living
organisms.
mRNA
Responsible for carrying
the genetic material to
the ribosomes and
insists as to what kind
of proteins is required
by the body.

9. Tools of Recombinan
DNA
Restriction Enzymes, Vector, Viruses & Bacteriophage

10. Tools of Recombinan DNA
• The enzymes which include the restriction endonucleases
• – help to cut DNA molecule from specific nitrogeneus base
(the polymerases)
• - help to synthesize DNA molecul and bind it (the ligases)
• They are two types, namely endonucleases and
exonucleases: The endonucleases is the restriction enzyme
which cut DNA molecule at interior side. The exonucleases
cut in exterior side of DNA and cut the nucleotides from the
ends of the DNA strands.
• The restriction endonucleases used in recombinant DNA
technology play a major role in determining the location at
which the desired gene is inserted into the vector genome.
Restriction
Enzymes
A recombinant DNA technology can be complete and achieved with the help of some elemental
tools. The different tools used for the purpose are: Restriction Enzymes, Vectors, Host organism.

11. ■ Restriction enzymes cut DNA molecules at specific locations
to produce restriction fragments with blunt ends or sticky
ends.
■ (a) Rsal restriction enzymes produce blunt-ended fragments
(b) EcoRI produces sticky ends by hanging at the 5 'end (c)
KpnI produces a sticky end by hanging at the 3' end.

12. • any DNA molecule that has the abylity to replicated inside the host which the
desiride gene has integrated for cloning
• Yeast cells, viruses, plasmids are the most commonly used vectors
• The vectors help in carrying and integrating the desired gene
• These form a very important part of the tools of recombinant DNA technology as
they are the ultimate vehicles that carry forward the desired gene into the host
organism
• To choose a suitable cloning vector for research, several things that must be
displayed are the size of the DNA fragment to be transferred
Cloning
Vectors
Tools of Recombinan DNA
■ Vector – DNA source which can replicate and is used to carry
foreign genes or DNA fragments.
■ Recombinant DNA – A vector that has taken up a foreign piece
of DNA.
A kind of host cell with contain vectors
are: transformed with alterated vector,
and with recombinan vector

13. ■ Overhanging sticky ends will complementarily
base pair, creating a recombinant DNA
molecule.
■ DNA ligase will seal the nick in the
phosphodiester backbone.
■ After cutting DNA with restriction enzymes, the
fragments can be separated on an agarose gel.
○ The smaller fragments will migrate
further than the longer fragments in an
electric field.
○ The bands are compared to standard
DNA of known sizes. This is often called
a DNA marker, or a DNA ladder.

14. • Host organism is the organism into which
the recombinant DNA is introduced.
• The host is the ultimate tool of
recombinant DNA technology which takes
in the vector engineered with the desired
DNA with the help of the enzymes.
• There are a number of ways in which this
recombinant DNA’s are inserted into the
host, namely – microinjection, biolistics or
gene gun, alternate cooling and heating,
use of calcium ions.
• Plants cells can also be transformed using
electroporation, which uses an electric
shock to make the cell membrane
permeable to plasmid DNA.
Competent
Host
Organism
Tools of Recombinan DNA
A gene gun uses biolistics to
insert DNA into plant tissue

15. STEP IN
RECOMBINAN
DNA, RNA, fungtion of RNA, Diferent kind of DNA vs RNA

16. •Getting gene interset from genomic library, cDMA
library, chemical syntesis of gene that knowing
subsquence
Identification and
isolation of gene
interest or DNA
fragment to be
cloned
•Cut plasmiid side by
Retriction enzyme
•Bind it by ligase
Introduction
suitable vector into
sutable organism or
host
(tranformation)
•Phiscal gene
tranfer
mehtode
•Chemical gene
transfer
methode
•DNA
imbibitions by
cell, tissues,
organs that call
Transformation
Insertion of
isolation gene into
suitable vector as
plasmid.
1
2&3
4

17. •3 kind of host cell: non
transformed,
transformed with
alterated vector, and
with recombinan vector.
•Can sellected by
antibiotic resisten in
selected medium, visible
characters, Assay for
biological activity, colony
hibriditation, blotting test
Selection the
transformened
host cell
Multiplication or
expression of
introduced gene
in the host
Analyse new
gene and protein
that pduce by
5
Cutting the gene at the recognition sites Polymerase chain reaction
Insertion of the desired recombinant DNA
into the host organism.

18. APPLICATION

19. Thank You