器官再生的新途径
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NheI
The blue letters represent the FRT sequence (48 bp).
The middle sequences represent KpnI, SalI, SacI and XhoI restriction enzyme sites, respectively.
1, without heat shock treatment
Southern blotting analysis of regenerated plants
Lanes 1–6, without heat shock treatment;
Lanes 1T–6T, heat shock treatment; WT, wild type plants; P, pLFFLPBBM plasmid as positive control. Radiolabelled BBM cDNA was used as a probe
RT-PCR of regenerated plants
Fig:BBM steady-state mRNA-levels
17
Separate a gene of interest from its promoter
Control of gene expression
Remove the selectable marker gene
35S-BBM-NOS
HSP18.2-FLP-NOS
FRT fusion sequences
EcoRI
5’GGGAATTCGAAGTTCCTATACTTTCTAGAGAATAGGA ACTTCGGAATAGGAACTTCGGTACCTATGTCGACGTAG AGCTCGACTCGAGGAAGTTCCTATACTTTCTAGAGAAT AGGAACTTCGATAGGAACTTCGCTAGCGG-3’
Technology prospects
Plant regeneration through tissue culture
organogenesis
somatic embryogenesis Binary Expression vector system
BABY BOOM gene(BBM)
FRT/FLP system
Heat shock-inducible system
3
Construction of gene cassettes
Plant transformation and culture
Heat shock induction
Main parts of the vector
The synthetic FRT sequence
Poplar regeneration via somatic embryogenesis
Transformation efficiency : 29%
Phenotypic characteristics of wild-type and transgenic plants
1T, heat shock treatment
7
Heat shock
Rooted cuttings
42℃ for 2h Two cycle Recovered at 21 ℃ for 12h
Incubated for 60d
8
Construction of fusion gene
Induction of plant regeneration via somatic embryogeห้องสมุดไป่ตู้esia
pBIN19
pLF
pBIN19+ FRT fusion sequences
pLFBBM
pLF + 35S-BBM-NOS
pLFFLPBBM
pLFBBM +HSP18.2-FLP-NOS
6
the calli of Chinese white poplar
Agrobacterium tumefaciens-mediated(LBA4404) WPM medium 5–6 weeks of culture somatic embryos were formed grew into plantlets The stem cuttings were inserted into oasis propagation tray
A novel method for induction of plant regeneration via somatic embryogenesis
From:Plant Science
Contents
2 3 4
Introduction Materials and Methods
Results and Discussion
The blue letters represent the FRT sequence (48 bp).
The middle sequences represent KpnI, SalI, SacI and XhoI restriction enzyme sites, respectively.
1, without heat shock treatment
Southern blotting analysis of regenerated plants
Lanes 1–6, without heat shock treatment;
Lanes 1T–6T, heat shock treatment; WT, wild type plants; P, pLFFLPBBM plasmid as positive control. Radiolabelled BBM cDNA was used as a probe
RT-PCR of regenerated plants
Fig:BBM steady-state mRNA-levels
17
Separate a gene of interest from its promoter
Control of gene expression
Remove the selectable marker gene
35S-BBM-NOS
HSP18.2-FLP-NOS
FRT fusion sequences
EcoRI
5’GGGAATTCGAAGTTCCTATACTTTCTAGAGAATAGGA ACTTCGGAATAGGAACTTCGGTACCTATGTCGACGTAG AGCTCGACTCGAGGAAGTTCCTATACTTTCTAGAGAAT AGGAACTTCGATAGGAACTTCGCTAGCGG-3’
Technology prospects
Plant regeneration through tissue culture
organogenesis
somatic embryogenesis Binary Expression vector system
BABY BOOM gene(BBM)
FRT/FLP system
Heat shock-inducible system
3
Construction of gene cassettes
Plant transformation and culture
Heat shock induction
Main parts of the vector
The synthetic FRT sequence
Poplar regeneration via somatic embryogenesis
Transformation efficiency : 29%
Phenotypic characteristics of wild-type and transgenic plants
1T, heat shock treatment
7
Heat shock
Rooted cuttings
42℃ for 2h Two cycle Recovered at 21 ℃ for 12h
Incubated for 60d
8
Construction of fusion gene
Induction of plant regeneration via somatic embryogeห้องสมุดไป่ตู้esia
pBIN19
pLF
pBIN19+ FRT fusion sequences
pLFBBM
pLF + 35S-BBM-NOS
pLFFLPBBM
pLFBBM +HSP18.2-FLP-NOS
6
the calli of Chinese white poplar
Agrobacterium tumefaciens-mediated(LBA4404) WPM medium 5–6 weeks of culture somatic embryos were formed grew into plantlets The stem cuttings were inserted into oasis propagation tray
A novel method for induction of plant regeneration via somatic embryogenesis
From:Plant Science
Contents
2 3 4
Introduction Materials and Methods
Results and Discussion