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Lecture 3 PowerPoint PPT Presentation

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Lecture 3. Ti plasmid derived vector system. Ti Plasmid. T-DNA region. DNA between L and R borders is transferred to plant as ssDNA; T-DNA encoded genes can be substituted by target genes. Tumor- producing genes. Opine catabolism. Virulence region. ORI. Ti-plasmid based vectors.

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Lecture 3

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Lecture 3

Ti plasmid derived vector system

Ti Plasmid



DNA between

L and R borders is

transferred to plant

as ssDNA;

T-DNA encoded genes can be substituted by target genes




Opine catabolism

Virulence region


Ti-plasmid based vectors

Binary systems

Co-integrated vectors

Needs 3 vectors

Needs 2 vectors:

Disarmed Ti plasmid

capable for infection

Disarmed Ti plasmid

with gene of interest

(no vir genes)

Helper vector

for infection

(with vir genes)


co-integrated plasmid

after homologous

recombination on T-DNA

Intermediate vector

with T-region

and gene of interest

(transferred by conjugation)

Helper vector

for transfer of

intermediate plasmid into A.tum

Co-integrated vectors (hybrid ti-plasmids)

Right now

rarely used


1) Long homologies required between the Ti plasmid

and the E. coli plasmids (pBR322 based Intermediate vectors)

making them difficult to engineer and use

2) Relatively inefficient gene transfer compared to the binary vecto

Ti plasmid vector systems are often working as binary vectors

T DNA region removed

Gene of interest

Plant selectable marker








Bacterial selectable


ori for A. tum

ori for A. tumefaciens

ori for E.coli

DISADVANTAGE: Depending on the orientation,

plasmids with two different origins of replication may be unstable in E. coli

ADVANTAGE: small vectors are used, which increases transfer efficiency

from E. coli to Agrobacterium.

No intermolecular recombination is needed

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