Plant Expression System
Download

Plant Expression System







Advertisement
/ 12 []
Download Presentation
Comments
gaetana
From:
|  
(115) |   (0) |   (0)
Views: 115 | Added: 20-02-2012
Rate Presentation: 2 0
Description:
Plant Expression System. M olecular B iology Lab oratory Oh Chang Jae. Overview of methods for plant transformation. A. Delivery of DNA into a single plant cell → transient assay B. Integration of DNA into the plant cell genome
Plant Expression System

An Image/Link below is provided (as is) to

Download Policy: Content on the Website is provided to you AS IS for your information and personal use only and may not be sold or licensed nor shared on other sites. SlideServe reserves the right to change this policy at anytime. While downloading, If for some reason you are not able to download a presentation, the publisher may have deleted the file from their server.











- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - E N D - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -




Slide 1

Plant Expression System

Molecular Biology Laboratory

Oh Chang Jae

Slide 2

Overview of methods for plant transformation

A. Delivery of DNA into a single plant cell→ transient assay

B. Integration of DNA into the plant cell genome

C. Regenerationof the transformed cell into a whole plant

Slide 3

In planta transformation with Agrobacterium

Vacuum infiltration

(Bechtold et al. 1993 )

Floral dipping method

(Bent et al., 1998)

Co-cultivation

with Agrobacterium

Slide 4

Agrobacterium tumefaciens

- crown gall disease

- Ti plasmid : T-DNA + vir genes

( > 200 Kbp )

Slide 5

Host Range

… cause disease primarily on dicots.

However, they can even infect gymnosperms,

not infectious on monocots, although transfer of the T-DNA into monocot can occur.

Pathogenesis

1. Tumor formation caused by the elevation of auxin and cytokinin levels by genes on a natural T-DNA

2. Opines synthesis

T-DNA used for scientific experimentation have usually been deleted of iaaM, iaaH, ipt and opine synthesis genes. → “ disarmed”

Slide 6

T-DNA transfer as mediated by vir gene functions

Slide 7

Plant transformation vector

Binary vector system

- Basic cloning vector (plant expression vector)

- plant transformation vector

Cointegrate vector system

- Cointegrate vector

- Disarmed Ti plasmid

Slide 8

General Vector composition

√T-DNA cassette

: Ti-plasmid right border (RB) + MCS + positive selectable marker + Ti-plasmid left border (LB)

√Promoter

Constitutve : 35S CaMV (Cauliflower mosaic virus)

Inducible : heat shock, glucocorticoid hormones, alcohol etc.

A plant promoter will often work in many different plant species, but yeast or human or bacterial promoter do not function.

√Terminator

Derived from CaMV, nopaline synthase gene etc.

√Common plant selectable marker : antibiotics , herbicides

Slide 9

Schematic diagram of binary vector

Slide 10

Basic cloning vector (pRT101)

P35S : CaMV 35S Promoter

pACaMV :CaMV polyadenylation site

ColE1 replication origin

bla ; Amp resistance gene

Slide 12

Plant transformation vector (pBIN20)

Kanamycine Resistance

NOS promoter

NOS terminator

Border


Copyright © 2014 SlideServe. All rights reserved | Powered By DigitalOfficePro