Restriction Analysis of pDRK and pGRN Original Plasmids

1
Restriction Analysis of pDRK and pGRNOriginal
Plasmids
ara C

H
i
n
d
I
I
I
ampr
PBAD
gfp
p
D
R
K
p
G
R
N
H
i
n
d
I
I
I
H
i
n
d
I
I
I
kanr
2
Restriction Analysis of pDRK and pGRNRestriction
Fragments after digest with HindIII
kanr
ara C
PBAD
ampr
gfp
3
Ligation of pDRK/pGRN Restriction Fragments
4
Ligation of pDRK/pGRN Restriction
FragmentsRecombinant Plasmid of Interest
5
Preparing Competent Cells for Transformation
6
Transforming Escherichia coli With a Recombinant
Plasmid
adhesion zone
Peptidoglycan layer
phospholipids
calcium ion
plasmid
7
GFP Expression
8
GFP Expression
ara C gene
gfp gene
promoter
C protein prevents gfp transcription by causing a
loop to form in the region of the gfp gene
9
GFP Expression
Arabinose C protein complex prevents DNA
looping and helps to align RNA polymerase on the
promoter site.
10
Transforming Escherichia coli With a Recombinant
Plasmid
Recombinant plasmids
transformed cells
Competent cells
11
Growth of transformed bacteria on various plates
P plates
LB LB/amp
LB/amp/ara
P- plates
No growth
LB LB/amp
12
GFP
13
Ligation of pDRK/pGRN Restriction
FragmentsRecombinant Plasmid of Interest
14
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