Introduction Components of binary vector Development of binary vector system Properties of binary vector Types of binary vector Plant transformation using binary vector Advantage of using binary vector Conclusion References

1. By
KAUSHAL KUMAR SAHU
Assistant Professor (Ad Hoc)
Department of Biotechnology
Govt. Digvijay Autonomous P. G. College
Raj-Nandgaon ( C. G. )

2. SYNOPSIS
 Introduction
 Components of binary vector
 Development of binary vector system
 Properties of binary vector
 Types of binary vector
 Plant transformation using binary vector
 Advantage of using binary vector
 Conclusion
 References

3. INTRODUCTION
 In these systems, the T-DNA region containing a gene of
interest is contained in one vector and the vir region is
located in a separate disarmed (without tumor-genes) Ti
plasmid. The plasmids co-reside in Agrobacterium and
remain independent.
 In the binary vector system, the two different plasmids
employed are:
 a wide-host-range small replicon, which has an origin of
replication (ori) that permits the maintenance of the plasmid
in a wide range of bacteria including E. coli and
Agrobacterium.
 a helper Ti plasmid, harbored in A. tumefaciens, which
lacks the entire T-DNA region but contains an intact vir
region.

5. COMPONENT OF BINARY VECTOR
 The standard component of binary vector are:
 Multiple cloning site
 A broad host range origin of replication functional in both
E.coli and A. tumefaciens.
 Selectable markers for both bacteria and plants
 T-DNA border sequences.

6. DEVELOPMENT OF BINARY VECTOR
 Hoekema et al. (1983) and de Framond et al.
(1983) determined that the vir and T-DNA regions of Ti-
plasmids could be split onto two separate replicons.

7. •Genes of interest are maintained within the T-DNA
region of a binary vector. Vir proteins encoded by genes
on a separate replicon (vir helper) mediate T-DNA
processing from the binary vector and T-DNAtransfer
from the bacterium to the host cell.
• The selection marker is used to indicate successful
plant transformation. ori, Origin of replication; Abr,
antibiotic-resistance gene used to select for the
presence of the T-DNAbinary vector in E.Coli
or in Agrobacterium.

8. PROPERTIES OF BINARY VECTOR
These include the following:
 T-DNA left and right border repeat sequences to define and
delimit T-DNA.
 A plant-active selectable marker gene .
 Restriction endonuclease, rare-cutting, or homing
endonuclease sites within T-DNA into which goi can be
inserted.
 Origin(s) of replication to allow maintenance in E. coli and
Agrobacterium.
 Antibiotic-resistance genes within the chromosome and
within backbone sequences for selection of the binary vector
in E. coli and Agrobacterium.

9. TYPES
 pGA series vectors
 pCG series vectors
 pCIT series
 Pgptv
 pBECK2000 series
 pGreen series

10. PLANT TRANSFORMATION USING
BINARY VECTOR
 Propagate binary vector in E. coli
 Isolate binary vector from E.coli and engineer (introduce a
foreign gene)
 Isolate engineered binary vector and introduce into
Agrobacterium containing a modified Ti plasmid
 Infect plant tissue with engineered Agrobacterium (T-DNA
containing the foreign gene gets inserted into a plant cell
genome)

11. Advantages of using Binary vector
 Binary vector do not need in vivo recombination.
 Binary vectors require only that an intact plasmid vector
to be introduced into the target bacterium, making the
process of bacterial transformation both more efficient
and quicker.
 Binary vector systems with plant ready genes in
Agrobacteria are easily and efficiently obtained.
 In binary system, the binary plasmids exist as separate
replicons, thus copy number is not strictly tied to that of
Ti plasmid.

12. CONCLUSION
 T-DNA binary systems have greatly simplified the generation
of transgenic plants. Along with companion vir helper
strains, numerous different T-DNA binary vectors with
specialized properties have been designed to facilitate such
diverse activities as protein expression, activation tagging,
protein localization, protein-protein interaction studies, and
RNAi-mediated gene silencing.

13. REFERENCES
 MK Razdan and SS Bhojwani- Plant tissue culture
 H.S. Chawla- Introduction to plant biotechnology
 IK Vasil- plant cell and Tissue culture