The document discusses plasmid and phage vectors used in genetic engineering, focusing on their properties, types, and applications in recombinant DNA technology. It describes plasmids as circular DNA molecules that can replicate independently and are used for gene cloning and expression, whereas bacteriophages, which infect bacteria, serve as vectors to deliver new genes. The conclusion highlights that while bacteriophages may damage their host, they provide effective tools for genetic manipulation compared to plasmids.

1. SHRI SHANKARACHARYA MAHAVIDYALAYA
Junwani ,BHILAI (C.G.)
PLASMID & PHAGE VECTOR
Dr. Rachana Choudhary
Dept. of Microbiology

2. SYNOPSIS
• INTRODUCTION
• VECTOR
• PROPERTIES OF VECTOR
• TYPES OF VECTOR
•PLASMIDAS VECTOR
• CLASSIFICATION
• TYPES OF PLASMIDS
• PROPERTIES OF PLASMID VECTOR
• USES OF PLASMID VECTOR
•BACTERIOPHAGE AS VECTOR
• TYPES OF BACTERIOPHAGE AS A
VECTOR
• λ PHAGE
• M13 PHAGE
• MACHENISM OF GENE CLONING
• APPLICATION
• CONCLUSION
• REFERENCE

3. INTRODUCTION
•Genetic engineering is the manipulation of genetic
material of an organism to give altered expresion of
our choice.
•It involves construction of recombinant DNA contain
DNA from different organisms.

4. PREPARATION
OF
RECOMBINANT
PLASMID
VECTOR

5. Introduction to Recombinant DNA Technology and
DNA Cloning
• Desired Gene (target gene)
• Restriction & Ligation
enzymes
• Vector
• Host

6. VECTOR
•Vectors are the DNA
molecules, which can carry a
foreign DNA fragment into a
host cell. .
•If it is used for the
reproducing the DNA
fragment, called cloning
vector.
•Also called vehicle DNA
because they act as carrier
of gene to be cloned into a
recipient cell.
•They are self replicating in
an appropriate host cell. Fig:- vectors

7. PROPERTIES OF VECTOR
•Small DNA molecules
•Origin of replication
•Unique restriction enzyme are able to replicate
autonomously
•Be non toxic to host cell
•Have space for foreign insert
•The vector shouldhave suitable marker genes that allow
easy selection of the transformed host cell.
•Unique recognition site

8. TYPES OF VECTOR
•There are three commonly used types of vectors:
1) plasmidvectors (e.g., pUC plasmids);
2) bacteriophage vectors (e.g., phage λ); and
3) phagemidvectors (e.g., pBlueScriptTM).

9. PLASMID vector
The term plasmid was first introduced by the American
molecular biologists Joshua Lederberg in 1952.
A plasmid is also known as replicon and extra
chromosomal DNA.
They are double stranded DNA molecule mostly occur in
circular forms.
They are naturally found in prokaryotes but some times found
in lower eukaryotes organisms.
Ex. Psc101,pBR322,& pUC18/19 etc.

10. a bacteriumshowing chromosomal DNA and plasmids. Not to
scale.

11. Classifications
Plasmids can be broadly classified into conjugative
plasmids and non-conjugative plasmids.
 Conjugative plasmids contain a set of transfer or tra
genes which promote sexual conjugation between
different cell.
 Non-conjugative plasmids are incapable of initiating
conjugation, hence they can be transferred only with
the assistance of conjugative plasmids.

12. Types Of Plasmids
1.Fertility [ F-plasmids ].
2. Resistance [ R-plasmids ].
3. Col plasmids.
4. Degradative plasmids.
5. Virulence plasmids.

13. PROPERTIES OF PLASMIDS
The size of plasmid ranges from 1 to 1000Kb.
Plasmid have their own origin of replication and
they have several unique restriction sites.
They can replicate independently of the host cells.
Plasmids have low molecular weight and high copy
number.
Easily isolated and integrated.
Used as a vectors.

14. pBR322
• The most versatile & cloning
vector in r DNA technology is
pBR-322
• Where, p-plasmid
• B- Boliver
• R-Rodrigver
• It carries genes resistance for
ampicillin & tetracycline
• Unique recognition sites for the
action of restriction endonuclease
 322- strain which is
distinguished from other
plasmids produced in the same
lab.

15. USES OF PLASMID VECTOR
1. Gene expression, i.e. production of gene products.
e.g. The first mammalian product produced in bacteria
was somatostatin (1977).
e.g. Gene expression using the pET system.
2. Gene cloning.
3. Cloning of complementary DNA (cDNA), i.e. DNA
synthesized using mRNA as a template and reverse
transcriptase.

16. BACTERIOPHAGE AS VECTOR
Bacteriophage
•A bacteriophage is a virus that
infects and replicates within
bacteria .
Structure of Bacteriophage
• Made of nucleoprotein
•ssDNA /sdDNA
•It consist head & tail.
Fig:- bacteriophage

17. 1. BACTERIOPHAGE LAMBDA
• Virus of E.coli
• Lytic or Lysogenic life style
• Lytic cycle
do not integrate their DNA into the host
bacterium genome and they usually kill
the host (lytic)
• Temperate phage may integrate into the
host genome causing lysogeny

18. LIFE CYCLE OF LAMBDA PHAGE
Fig:- Diagramatic representation of Life cycle of Lambda phage.

19. LAMBDA PHAGE AS VECTOR
•Manipulation of restrictio site
•Size limitation for packaging
•Transfection of recombinant molecules
The problem of transfection of recombinant molecules constructed
in vitro was overcome by the successful in vitro assembly of viable
and infectious phage particles.
• two strain packaging
• single strain packaging

20. DNAcloning using lambda phage as vector

21. Fig:- Generalized transuding phage life cycle.
Phage adsorption
to donor Phage DNA injection
Phage DNA replication
Phage Lytic Cycle
Host lysis
Virion assembly
Transducing particle
adsorption to recipient
Donor DNA
injection
Homologous recombination
with recipient DNA
Transductant

22. M13 PHAGE
Introduction
•M13 phage is a filamentous
bacteriophage with single
stranded DNA genomes 6400
bases in length.
•they require the E.coli F pilus
for infection.
•900 nm long and 6-7 nm in
diameter
Fig:- M13 bacteriophage

23. Genome structure of M13 phage

24. LIFE CYCLE OF M13 PHAGE

25. Fig:- bacteriophage M13 cloning system

26. APPLICATION OF PHAGE
• Cloning
• Ss DNA for probes, sequencing
• Phage display technology, M13 will produce foreign
protein on surface as part of its protein coat, can use to
generate specific antibodies
•Cloning pieces too big for plasmids
•Generation of ds DNA

27. CONCLUSION
•Bacteriophages are good vector as compare to plasmid
•A bacteriophage vector is a virus that infects a
bacterium and inserts its DNA or RNA into the host to
provide new genes coding for new proteins.
•However, bacteriophages frequently destroy its host
whereas plasmids don't.

28. Reference
•Biotechnology by B.D. Singh
•Genetics by P.K. Gupta
•Genetics by T.A. Brown
•Biotechnology by U. Satyanarayana
•Google Search

29. THANKS