for pharmacy and medical student..

1. DEPARTMENT OF PHARMACY,
Dr. BHIMRAO AMBEDKAR UNIVERSITY,
AGRA- 282002
Submitted By: Submitted To:
Rohit Yadav Dr. R. S. Sharma
B.Pharma (Asst. Professor)
VIII- SEM
Topic – Genetic Recombination

2. Contents
Working Glossary
Transformation,
conjugation,
transduction,
 protoplast fusion.
Gene cloning and their applications.
Development of hybn'doma for mono clonal
antibodies.
Study of drugs produced by biotechnology
such as Actives, Humulin,Humatrope, and HB
etc.

3. INTRODUCTION
Genetic recombination is the process by which genetic
elements from two separate sources are brought together
in a single unit.
At molecular level, recombination can be though to as the
movement of genetic information from one molecule of
nucleic acid to another.
The genetic exchange occurring between homologous
DNA sequences from two different sources is termed as
general recombination. For this to happen, identical
sequences on the two recombining molecules are required.
The process of genetic exchange which occurs in
eukaryotes during sexual reproduction(meiosis)is an
example of this type of genetic recombination.

4. Auxotroph: A mutated microorganism having nutritional
requirements that differ from those of unmuted microorganisms
from the same strain.
Cloning vector: Genetic element into which genes can be
recombined and replicated.
Conjugation: Transfer of genes from one prokaryotic cell to
another by a mechanism involving cell-to-cell contact and a
plasmid.
Diploid: A eukaryotic cell or organism containing two sets of
chromosomes.
Electroporation: The use of an electric pulse to induce cells to
take up free DNA .
Gene disruption: Use of genetic techniques to inactivate a gene
by inserting within it a DNA fragment containing an easily
selectable marker. The inserted fragment is called a cassette, and
the process of insertion, cassette mutagenesis.

5. Genetic map: The arrangement of genes on a chromosome.
Genome: the total complement of genes of a cell or a virus.
Genotype: The precise genetic makeup of an organism.
Hybridization: formation of a duplex nucleic acid molecule with strands
derived from different sources by complementary base pairing Molecular
cloning: Isolation and incorporation of a fragment of DNA into a vector
where it can be replicated.
Haploid: A cell or organism that has only one set of chromosomes.
Mutagens: Agents that cause mutation.
Mutant: An organism whose genome carries a mutation.
Mutation: An inheritable change in the base sequence of the genome of an
organism.

6. Nucleic acid probe: a strand of nucleic acid that can be labeled and used to
hybridize to a complementary molecule from mixture of other nucleic acids
Phenotype: the observable characteristics of an organism.
 Plasmid: an extra chromosomal genetic element that has no extracellular
form Point.
Polymerase chain reaction (PCR): a method used to amplify a specific
DNA sequence in vitro by repeated cycles of synthesis using specific
primers and DNA polymerase.
Recombination: the process by which parts or all of the DNA molecules
from two separate sources are exchanged or brought together into a single
unit.

7. Restriction enzyme: an enzyme that recognizes and makes double-stranded
breaks at specific DNA sequences Shotgun.
Cloning: making a gene library by closing random DNA fragments Site-
directed.
Mutagenesis: a technique whereby a gene with a specific mutation can be
constructed in vitro Synthetic.
DNA: a DNA molecule made by a chemical process in a laboratory.
Transduction: transfer of host genes from one cell to another by a virus.
Transformation: transfer of bacterial genes involving free DNA.

8. TRANSFORMATION
In molecular biology, transformation is the genetic alteration of
a cell resulting from the direct uptake and incorporation of exogenous
genetic material from its surroundings through the cell membrane.
Transformation" may also be used to describe the insertion of new genetic
material into nonbacterial cells, including animal and plant cells; however,
because "transformation" has a special meaning in relation to animal cells,
indicating progression to a cancerous state, the process is usually called
"transfection
HISTORY
Transformation in bacteria was first demonstrated in 1928 by the British
bacteriologist Frederick Griffith. Griffith was interested in determining
whether injections of heat-killed bacteria could be used to vaccinate mice
against pneumonia. However, he discovered that a non-virulent strain
of Streptococcus pneumoniae could be made virulent after being exposed to
heat-killed virulent strains. Griffith hypothesized that some "transforming
principle" from the heat-killed strain was responsible for making the
harmless strain virulent. In 1944 this "transforming principle" was identified
as being genetic by Oswald Avery

10. *Bacterial transformation is used: To make
multiple copies of DNA, called DNA cloning. To
make large amounts of specific human proteins,
for example, human insulin, which can be used
to treat people with Type I diabetes. To
genetically modify a bacterium or other cell.

11. Bacterial conjugation is the transfer of genetic material between bacterial
cells by direct cell-to-cell contact or by a bridge-like connection between
two cells. This takes place through a pilus
History:-
*The process was discovered in 1946 by Joshua Lederberg and Edward
Tatum.
MACHANISM
Conjugation diagram:-
Donor cell produces pilus.
Pilus attaches to recipient cell and brings the two cells together.
The mobile plasmid is nicked and a single strand of DNA is then
transferred to the recipient cell.
Both cells synthesize a complementary strand to produce a double stranded
circular plasmid and also reproduce pili; both cells are now viable donor for
the F-factor

13. Transduction is the process by which foreign DNA is introduced into a
cell by a virus or viral vector.An example is the viral transfer
of DNA from one bacterium to another and hence an example
of horizontal gene transfer.Transduction does not require physical contact
between the cell donating the DNA and the cell receiving the DNA.
Discovery: Transduction was discovered by Norton
Zinder and Joshua Lederberg in 1952 in Salmonella.
Type of Transduction:-
1. Generalized transduction.
2. Specialized transduction.

14. Generalized transduction:- Generalized transduction is the process
by which any bacterial DNA may be transferred to another bacterium via a
bacteriophage. It is a rare event; a very small percentage of phage particles
happen to carry a donor bacterium's DNA.
Specialized transduction:- Specialized transduction is the process
by which a restricted set of bacterial genes is transferred to another
bacterium. The genes that get transferred (donor genes) depend on where the
phage genome is located on the chromosome.
Medical applications:-
*Resistance to antibiotic drugs.
*Gene therapy: Correcting genetic diseases by direct
modification of genetic errors

17. Protoplast fusion, also called Somatic fusion is a type of genetic
modification in plants by which two distinct species of plants are fused
together to form a new hybrid plant with the characteristics of both,
a somatic hybrid. Hybrids have been produced either between different
varieties of the same species (e.g. between non-flowering potato plants
and flowering potato plants) or between two different species (e.g.
between wheat Triticum and rye Secale to produce Triticale.
Process for plant cells:-
*The somatic fusion process occurs in four steps:
1. The removal of the cell wall of one cell of each type of plant
using cellulase enzyme to produce a somatic cell called a protoplast
2. The cells are then fused using electric shock (electrofusion) or
chemical treatment to join the cells and fuse together the nuclei.
The resulting fused nucleus is called heterokaryon.
3. The formation of the cell wall is then induced using hormones
4. The cells are then grown into calluses which then are further grown
to plantlets and finally to a full plant, known as a somatic hybrid.

18. Somatic cells of different types can be fused to obtain hybrid cells. Hybrid
cells are useful in a variety of ways, e.g.,
(i) to study the control of cell division and gene expression,
(ii) to investigate malignant transformations,
(iii) to obtain viral replication.
(iv) for gene or chromosome mapping.
(v) production of monoclonal antibodies by producing hybridoma (hybrid cells
between an immortalised cell and an antibody producing lymphocyte), etc.
Chromosome mapping through somatic cell hybridization is essentially based
on fusion of human and mouse somatic cells. Generally,
human fibrocytes or leucocytes are fused with mouse continuous cell lines

19. * Gene cloning (DNA cloning) The production of exact copies (clones) of
a particular gene or DNA sequence using genetic engineering techniques.
* The DNA containing the target gene is split into fragments
using restriction enzymes. These fragments are then inserted into
cloning vectors.
* such as bacterial plasmids or bacteriophages, which transfer the
recombinant DNA to suitable host cells, such as the bacterium E. coli.
Alternatively, complementary DNA is inserted into the vectorsInside the
host cell the recombinant DNA undergoes replication; thus, a bacterial
host will give rise to a colony of cells containing the cloned target gene.
* A clone is an exact copy of an organism, organ, single cell, or
macromolecule, clones can be used in many areas of biomedical and
industrial research. Genetic engineering is the process of cloning genes
into new organisms, or altering a genetic sequence to change the protein
product.

21. If You Want This Power point Presentation, Go to my slide share
account link given Below:
https://www.slideshare.net/RohitRishi4