Лекция_4_Б
.pdf
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WT6803 |
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WT6803 |
WT6803 |
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WT6803 |
WT6803 |
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licBM2/control licBM2/pilA |
licBM2/2016 licBM2/art (+) licBM2/art (-) |
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Agrobacterium tumefaciens:
/ #
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$ "
# " / $ " !""
T-DNA # !
(# )
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X " T-DNAs ; "
X. # "
Biolistics / Gene gun:
/ # "
Менее зависит от
генотипа
Можно использовать линейный фрагмент ДНК;
Вектора более простые
Высокая степень ко-
трансформации, но
возможна сегрегация
маркерных генов
X Множественные
Molecular Characterisation
Agrobacterium
1-5 "
0 !
30-40%
" #
"
SwaI or BglII gusA. probe
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Intron |
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Bgl II |
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Swa I |
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Bgl II |
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Nos 3’ |
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Bgl II |
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Swa I |
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RB |
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LB |
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Ubi 5’ + I |
Ubi 5’ + I Bar (PAT) |
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UidA(GUS) |
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NptI |
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Biolistics (whole plasmids or frags)
1 - >25 "
~5-10% " # ) # !) ) !
HpaI / bar probe
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DNA |
DNA |
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particle bombardment |
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Agrobacterium Ti plasmid mediation |
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particle bombardment |
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polyethylene glycol treatment |
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polyethylene glycol treatment |
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electroporation |
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nucleus
chloroplast
7
pLD2000-mALS
rbcL |
Prrn |
aadA |
T |
PpsbA |
mALS |
T |
accD |
rbcL |
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accD |
(
(
rbcL |
Prrn |
aadA |
T |
PpsbA |
mALS |
T |
accD |
(
rbcL: large subunit of Rubisco
aadA: aminoglycosid-3’-adenyl-transferase rrn: 16 S rRNA
psbA: photosystem II reaction center D1 protein accD: carboxytransfetase β sununit
7
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pLD2000-mALS |
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rbcL |
Prrn |
aadA |
T |
PpsbA |
mALS |
T |
accD |
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PCR analysis
ALS activity
spectinomycin (SP)- |
transplastomic |
resistance |
tobacco |
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(
(-. )
/
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