The production of antimicrobial polypeptides in chloroplasts:
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| Main Author: | |
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| Format: | Thesis Book |
| Language: | English |
| Published: |
Potsdam
August 2020
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| Subjects: | |
| Online Access: | Inhaltsverzeichnis Inhaltsverzeichnis |
| Physical Description: | xiii, 114 Seiten Illustrationen 30 cm |
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| 100 | 1 | |a Hoelscher, Matthijs Pieter |e Verfasser |0 (DE-588)1219005088 |4 aut | |
| 245 | 1 | 0 | |a The production of antimicrobial polypeptides in chloroplasts |c Matthijs Pieter Hoelscher |
| 264 | 1 | |a Potsdam |c August 2020 | |
| 300 | |a xiii, 114 Seiten |b Illustrationen |c 30 cm | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
| 338 | |b nc |2 rdacarrier | ||
| 502 | |b Dissertation |c Universität Potsdam |d 2020 | ||
| 655 | 7 | |0 (DE-588)4113937-9 |a Hochschulschrift |2 gnd-content | |
| 856 | 4 | 2 | |m B:DE-101 |q application/pdf |u https://d-nb.info/1221382128/04 |3 Inhaltsverzeichnis |
| 856 | 4 | 2 | |m DNB Datenaustausch |q application/pdf |u http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=032730632&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |3 Inhaltsverzeichnis |
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| adam_text | CONTENTS
E
IDESSTATTUCHE
E
RKLARUNG
..................................................................................................................................................
VI
D
ECLARATION
OF
CONTRIBUTIONS
....................................................................................................................
VII
A
BSTRACT
..................................................................................................................................................................................
VM
L
IST
OF
F
IGURES
AND
TABLES
....................................................................................................................................................
IX
A
BBREVIATIONS
.........................................................................................................................................................................
XI
1
GENERAL
INTRODUCTION
1
2
HIGH-LEVEL
EXPRESSION
OF
THE
HIV
ENTRY
INHIBITOR
GRIFFITHSIN
FROM
THE
PLASTID
GENOME
AND
RETENTION
OF
BIOLOGICAL
ACTIVITY
IN
DRIED
TOBACCO
LEAVES
............................
2
2.1
A
BSTRACT
.......................................................................................................................................
2
2.1.1
KEY
MESSAGE
.................................................................................................................................................................
2
2.1.2
A
BSTRACT
.....................................................................................................................................
2
2.2
I
NTRODUCTION
.................................................................................................................................
2
2.3
M
ATERIALS
AND
METHODS
................................................................................................................
4
2.3.1
P
LANT
MATERIAL
AND
GROWTH
CONDITIONS
......................................................................................
4
2.3.2
C
LONING
PROCEDURES
....................................................................................................................
5
2.3.3
C
HLOROPLAST
TRANSFORMATION
......................................................................................................
5
2.3.4
S
ELECTABLE
MARKER
GENE
REMOVAL
.................................................................................................
5
2.3.5
I
SOLATION
OF
NUCLEIC
ACIDS
AND
GEL
BLOT
ANALYSES
.........................................................................
5
2.3.6
P
ROTEIN
ISOLATION
,
PURIFICATION
AND
VISUALIZATION
........................................................................
6
2.3.7
D
ETERMINATION
OF
ANTI
-HIV
ACTIVITY
OF
PURIFIED
GRIFFITHSIN
..........................................................
7
2.4
R
ESULTS
........................................................................................................................................
11
2.4.1
STABLE
TRANSFORMATION
OF
THE
TOBACCO
PLASTID
GENOME
WITH
A
GRIFFITHSIN
EXPRESSION
CASSETTE
..11
2.4.2
S
ELECTABLE
MARKER
GENE
REMOVAL
FROM
TRANSPLASTOMIC
PLANTS
..................................................
11
2.4.3
G
RIFFITHSIN
M
RNA
AND
PROTEIN
ACCUMULATION
IN
TRANSPLASTOMIC
PLANTS
.....................................
12
2.4.4
P
URIFICATION
OF
GRIFFITHSIN
FROM
TRANSPLASTOMIC
TOBACCO
PLANTS
..............................................
14
2.4.5
A
NTI
-HIV
ACTIVITY
OF
CHLOROPLAST
-
EXPRESSED
GRIFFITHSIN
.............................................................
15
2.4.6
P
URIFICATION
OF
ACTIVE
GRIFFITHSIN
FROM
DRIED
TOBACCO
...............................................................
16
2.5
D
ISCUSSION
.................................................................................................................
16
2.6
A
CKNOWLEDGEMENTS
.....................................................................................................................
17
3
STRATEGIES
FOR
ANTIMICROBIAL
PEPTIDE
PRODUCTION
IN
CHLOROPLASTS
..........................
18
3.1
I
NTRODUCTION
...............................................................................................................................
18
3.2
M
ATERIALS
AND
METHODS
..............................................................................................................
25
3.2.1
DNA
DESIGN
OF
FUSED
ANTIMICROBIAL
PEPTIDES
..................................................................................................
25
3.2.2
G
ENERATION
OF
PLASTID
TRANSFORMATION
CONSTRUCTS
...................................................................
25
3.2.3
C
ONSTITUTIVE
HIGH
EXPRESSION
.....................................................................................................
26
3.2.4
C
HLOROPLAST
CONSTRUCTS
FOR
INDUCIBLE
EXPRESSION
.....................................................................
26
3.2.5
N
UCLEAR
ENCODED
,
ETHANOL
-
INDUCIBLE
,
PLASTID
TARGETED
T7
RNA
POLYMERASE
(P
ET
OH::
PT
T7RP)....26
3.2.6
P
REDICTION
OF
PROTEIN
PROPERTIES
BASED
ON
AMINO
ACID
SEQUENCE
................................................
28
3.2.7
P
ATHOGEN
INDUCIBLE
PLASTID
TARGETED
T7
RNA
POLYMERASE
..........................................................
28
3.2.8
DNA
PREPARATION
FOR
CHLOROPLAST
TRANSFORMATION
...................................................................................
29
3.2.9
C
HLOROPLAST
TRANSFORMATION
.....................................................................................................
29
3.2.10
S
OUTHERN
BLOTTO
CONFIRM
THE
HOMOPLASMIC
STATE
...................................................................
30
3.2.11
P
LANT
GROWTH
UNDER
STERILE
CONDITIONS
...................................................................................
30
3.2.12
S
EED
STERILIZATION
.....................................................................................................................
30
3.2.13
A
NTIBIOTIC
SELECTION
OF
SEEDLINGS
..............................................................................................
30
3.2.14
PLANT
GROWTH
IN
GREENHOUSE
...........................................................................................................................
30
3.2.15
G
RAFTING
..................................................................................................................................
30
3.2.16
P
EPTIDE
SYNTHESIS
......................................................................................................................
31
3.2.17
P
HENOL
BASED
PROTEIN
EXTRACTION
..............................................................................................
31
3.2.18
E
XTRACTION
PROCEDURE
TO
TEST
BUFFERS
FOR
F
AMP
EXTRACTION
....................................................
31
3.2.19
P
ROTEIN
EXTRACTION
BUFFERS
......................................................................................................
31
3.2.20
U
REA
EXTRACTION
BUFFERS
...........................................................................................................
31
3.2.21
T
RIS
-T
RICINE
SAMPLE
BUFFER
........................................................................................................
31
3.2.22
D
IRECT
EXTRACTION
WITH
T
RIS
-T
RICINE
SAMPLE
BUFFER
...................................................................
32
3.2.23
H
IS
-
TAG
PURIFICATION
FROM
PLANT
MATERIAL
................................................................................
32
3.2.24
T
RIS
-T
RICINE
SDS
PAGE
..............................................................................................................
32
3.2.25
COOMASSIE
STAINING
GELS
.....................................................................................................................................
33
3.2.26
W
ESTERN
BLOT
...........................................................................................................................
33
3.2.27
COOMASSIE
STAINING
OF
WESTERN
BLOT
...............................................................................................................
33
3.2.28
I
MMUNODETECTION
OF
WESTERN
BLOT
...........................................................................................
33
3.2.29
B
LUE
N
ATIVE
PAGE
.....................................................................................................................
33
3.2.30
T
HEOPHYLLINE
INDUCTION
OF
RA
MP
ER
PLANTS
...............................................................................
34
3.2.31
E
THANOL
INDUCTION
...................................................................................................................
34
3.2.32
T
ESTING
ANTI
-
MICROBIAL
ACTIVITY
IN
VITRO
........................................................................................
35
3.2.33
T
RANSIENT
TRANSFORMATION
OF
N
ICOTIANA
TABACUM
PLANTS
.........................................................
35
3.2.34
A
GROBACTERIUM
TRANSFORMATION
..............................................................................................
35
3.2.35
S
TABLE
TRANSFORMATION
OF
N
ICOTIANA
TABACUM
PLANTS
..............................................................
36
3.2.36
P
LANT
RESISTANCE
ASSAYS
PATHOGEN
INFECTION
.............................................................................
36
3.2.37
R
ECAPTURE
OF
BACTERIA
FROM
INFECTED
PLANTS
.............................................................................
37
3.2.38
SUMO
DIGESTION
....................................................................................................................................................
37
3.2.39
T
ESTING
CHLOROPLAST
TRANSFORMATION
CONSTRUCTS
IN
E
.
COLI
.........................................................
37
3.2.40
C
ONFOCAL
MICROSCOPY
...............................................................................................................
38
3.2.41
T
RANSMISSION
ELECTRON
MICROSCOPY
(TEM)................................................................................
38
3.3
R
ESULTS
.........................................................................................................................................
39
3.3.1
G
ENERATION
AND
VALIDATION
OF
TRANSPLASTOMIC
PLANTS
THAT
CONSTITUTIVELY
PRODUCE
F
AMP
S
.......
39
3.3.2
P
LANT
PHENOTYPES
CAUSED
BY
STRONG
CONSTITUTIVE
EXPRESSION
OF
F
AMP
GENES
..............................
43
3.3.3
F
AMP
ACCUMULATION
IN
CONSTITUTIVELY
EXPRESSING
PLANTS
............................................................
43
3.3.4
T
HE
INDUCIBLE
RA
MP
ER
SYSTEM
.....................................................................................................
44
3.3.5
P
LANT
PHENOTYPES
OF
RA
MP
ER::
F
AMP
PLANTS
..............................................................................
49
3.3.6
F
AMP
BACKGROUND
LEVELS
IN
RA
MP
ER
AND
P
T
7
PLANTS
..................................................................
50
3.3.7
RA
MP
ER
INDUCTION
OF
F
AMP
PRODUCTION
BY
THEOPHYLLINE
.........................................................................
50
3.3.8
T
HE
EFFECT
OF
THEOPHYLLINE
ON
PLANT
PHENOTYPES
.........................................................................
51
3.3.9
T
HE
EFFECT
OF
LEAF
AGE
ON
ACCUMULATION
.....................................................................................
51
3.3.10
F
AMP
EXTRACTION
UNDER
NATIVE
CONDITIONS
...............................................................................
51
3.3.11
T
HE
EFFECT
OF
F
AMP
ACCUMULATION
ON
CHLOROPLAST
INTEGRITY
....................................................
56
3.3.12
E
THANOL
-
INDUCIBLE
F
AMP
PRODUCTION
.......................................................................................
56
3.3.13
F
AMP
EXTRACTION
WITH
DIFFERENT
BUFFERS
AND
DETERGENTS
.........................................................
62
3.3.14
D
ENATURING
EXTRACTION
OF
3-N
OVESCANDIN
-5
............................................................................
64
3.3.15
T
HE
EFFECT
OF
DETERGENTS
,
DENATURING
AND
ACETIC
ACID
ON
THE
EXTRACTION
OF
3-NOVESCANDIN-5..64
3.3.16
H
IS
-
TAG
PURIFICATION
.................................................................................................................
65
3.3.17
I
N
VITRO
ANTIMICROBIAL
ACTIVITY
OF
F
AMP
S
..................................................................................
65
IV
3.3.18
S
USCEPTIBILITY
OF
F
AMP
PRODUCING
RA
MP
ER
PLANTS
TO
PATHOGENS
.............................................
67
3.3.19
P
ATHOGEN
INDUCIBLE
F
AMP
PRODUCTION
....................................................................................
72
3.3.20
SUMO
FUSIONS
FOR
ENHANCED
SOLUBILITY
AND
REDUCED
TOXICITY
...............................................................
76
3.3.21
A
CCUMULATION
OF
SUMO
FUSIONS
.............................................................................................
77
3.3.22
E
XTRACTION
OF
SUMO-3-W
AMDEPOSIN
-5
...................................................................................
78
3.3.23
SUMO
REMOVAL
....................................................................................................................................................
78
3.3.24
E
THANOL
-
INDUCIBLE
SUMO-3-W
AMDEPOSIN
-5
............................................................................
82
3.3.25
SUMO-HA-WAM1
TESTING
IN
E.
COLI
..............................................................................................................
82
3.3.26
C
LEAVAGE
OF
SUMO-HA-WAM1
FROM
E.
COU
................................................................................
82
3.4
D
ISCUSSION
...................................................................................................................................
83
3.4.1
F
AMP
PRODUCTION
IN
CHLOROPLASTS
............................................................................................
83
3.4.2
T
HE
INDUCIBLE
RA
MP
ER
SYSTEM
FOR
F
AMP
PRODUCTION
.................................................................
86
3.4.3
L
EAKY
ACTIVATION
OF
THE
T7
PROMOTER
.........................................................................................
86
3.4.4
RA
MP
ER
INDUCTION
OF
F
AMP
PRODUCTION
...................................................................................
87
3.4.5
E
T
OH
INDUCIBLE
EXPRESSION
OF
F
AMP
S
.........................................................................................
88
3.4.6
E
XTRACTION
AND
PURIFICATION
OF
F
AMP
S
......................................................................................
89
3.4.7
I
N
VITRO
ANTIMICROBIAL
ACTIVITY
OF
F
AMP
S
...................................................................................
90
3.4.8
D
ISEASE
RESISTANCE
OF
F
AMP
PRODUCING
PLANTS
...........................................................................
90
3.4.9
SUMO
AS
CARRIER
PROTEIN
FOR
(
F
)AMP
PRODUCTION
......................................................................................
92
3.5
C
ONCLUSION
..................................................................................................................................
95
4
GENERAL
DISCUSSION
96
R
EFERENCES
...........................................................................................................................................
98
SUPPLEMENTARY
FIGURES
....................................................................................................................................................
105
ALLGEMEINVERSTANDLICHE
ZUSAMMENFASSUNG
IN
DEUTSCHER
SPRACHE
..................................................................
109
CURRICULUM
VITAE
................................................................................................................................................................
110
P
UBLICATIONS
......................................................................................................................................
112
C
ONFERENCE
ATTENDANCE
......................................................................................................................
112
S
CHOLARSHIPS
AND
PRIZES
......................................................................................................................
112
A
CKNOWLEDGEMENTS
...........................................................................................................................
113
V
|
| adam_txt |
CONTENTS
E
IDESSTATTUCHE
E
RKLARUNG
.
VI
D
ECLARATION
OF
CONTRIBUTIONS
.
VII
A
BSTRACT
.
VM
L
IST
OF
F
IGURES
AND
TABLES
.
IX
A
BBREVIATIONS
.
XI
1
GENERAL
INTRODUCTION
1
2
HIGH-LEVEL
EXPRESSION
OF
THE
HIV
ENTRY
INHIBITOR
GRIFFITHSIN
FROM
THE
PLASTID
GENOME
AND
RETENTION
OF
BIOLOGICAL
ACTIVITY
IN
DRIED
TOBACCO
LEAVES
.
2
2.1
A
BSTRACT
.
2
2.1.1
KEY
MESSAGE
.
2
2.1.2
A
BSTRACT
.
2
2.2
I
NTRODUCTION
.
2
2.3
M
ATERIALS
AND
METHODS
.
4
2.3.1
P
LANT
MATERIAL
AND
GROWTH
CONDITIONS
.
4
2.3.2
C
LONING
PROCEDURES
.
5
2.3.3
C
HLOROPLAST
TRANSFORMATION
.
5
2.3.4
S
ELECTABLE
MARKER
GENE
REMOVAL
.
5
2.3.5
I
SOLATION
OF
NUCLEIC
ACIDS
AND
GEL
BLOT
ANALYSES
.
5
2.3.6
P
ROTEIN
ISOLATION
,
PURIFICATION
AND
VISUALIZATION
.
6
2.3.7
D
ETERMINATION
OF
ANTI
-HIV
ACTIVITY
OF
PURIFIED
GRIFFITHSIN
.
7
2.4
R
ESULTS
.
11
2.4.1
STABLE
TRANSFORMATION
OF
THE
TOBACCO
PLASTID
GENOME
WITH
A
GRIFFITHSIN
EXPRESSION
CASSETTE
.11
2.4.2
S
ELECTABLE
MARKER
GENE
REMOVAL
FROM
TRANSPLASTOMIC
PLANTS
.
11
2.4.3
G
RIFFITHSIN
M
RNA
AND
PROTEIN
ACCUMULATION
IN
TRANSPLASTOMIC
PLANTS
.
12
2.4.4
P
URIFICATION
OF
GRIFFITHSIN
FROM
TRANSPLASTOMIC
TOBACCO
PLANTS
.
14
2.4.5
A
NTI
-HIV
ACTIVITY
OF
CHLOROPLAST
-
EXPRESSED
GRIFFITHSIN
.
15
2.4.6
P
URIFICATION
OF
ACTIVE
GRIFFITHSIN
FROM
DRIED
TOBACCO
.
16
2.5
D
ISCUSSION
.
16
2.6
A
CKNOWLEDGEMENTS
.
17
3
STRATEGIES
FOR
ANTIMICROBIAL
PEPTIDE
PRODUCTION
IN
CHLOROPLASTS
.
18
3.1
I
NTRODUCTION
.
18
3.2
M
ATERIALS
AND
METHODS
.
25
3.2.1
DNA
DESIGN
OF
FUSED
ANTIMICROBIAL
PEPTIDES
.
25
3.2.2
G
ENERATION
OF
PLASTID
TRANSFORMATION
CONSTRUCTS
.
25
3.2.3
C
ONSTITUTIVE
HIGH
EXPRESSION
.
26
3.2.4
C
HLOROPLAST
CONSTRUCTS
FOR
INDUCIBLE
EXPRESSION
.
26
3.2.5
N
UCLEAR
ENCODED
,
ETHANOL
-
INDUCIBLE
,
PLASTID
TARGETED
T7
RNA
POLYMERASE
(P
ET
OH::
PT
T7RP).26
3.2.6
P
REDICTION
OF
PROTEIN
PROPERTIES
BASED
ON
AMINO
ACID
SEQUENCE
.
28
3.2.7
P
ATHOGEN
INDUCIBLE
PLASTID
TARGETED
T7
RNA
POLYMERASE
.
28
3.2.8
DNA
PREPARATION
FOR
CHLOROPLAST
TRANSFORMATION
.
29
3.2.9
C
HLOROPLAST
TRANSFORMATION
.
29
3.2.10
S
OUTHERN
BLOTTO
CONFIRM
THE
HOMOPLASMIC
STATE
.
30
3.2.11
P
LANT
GROWTH
UNDER
STERILE
CONDITIONS
.
30
3.2.12
S
EED
STERILIZATION
.
30
3.2.13
A
NTIBIOTIC
SELECTION
OF
SEEDLINGS
.
30
3.2.14
PLANT
GROWTH
IN
GREENHOUSE
.
30
3.2.15
G
RAFTING
.
30
3.2.16
P
EPTIDE
SYNTHESIS
.
31
3.2.17
P
HENOL
BASED
PROTEIN
EXTRACTION
.
31
3.2.18
E
XTRACTION
PROCEDURE
TO
TEST
BUFFERS
FOR
F
AMP
EXTRACTION
.
31
3.2.19
P
ROTEIN
EXTRACTION
BUFFERS
.
31
3.2.20
U
REA
EXTRACTION
BUFFERS
.
31
3.2.21
T
RIS
-T
RICINE
SAMPLE
BUFFER
.
31
3.2.22
D
IRECT
EXTRACTION
WITH
T
RIS
-T
RICINE
SAMPLE
BUFFER
.
32
3.2.23
H
IS
-
TAG
PURIFICATION
FROM
PLANT
MATERIAL
.
32
3.2.24
T
RIS
-T
RICINE
SDS
PAGE
.
32
3.2.25
COOMASSIE
STAINING
GELS
.
33
3.2.26
W
ESTERN
BLOT
.
33
3.2.27
COOMASSIE
STAINING
OF
WESTERN
BLOT
.
33
3.2.28
I
MMUNODETECTION
OF
WESTERN
BLOT
.
33
3.2.29
B
LUE
N
ATIVE
PAGE
.
33
3.2.30
T
HEOPHYLLINE
INDUCTION
OF
RA
MP
ER
PLANTS
.
34
3.2.31
E
THANOL
INDUCTION
.
34
3.2.32
T
ESTING
ANTI
-
MICROBIAL
ACTIVITY
IN
VITRO
.
35
3.2.33
T
RANSIENT
TRANSFORMATION
OF
N
ICOTIANA
TABACUM
PLANTS
.
35
3.2.34
A
GROBACTERIUM
TRANSFORMATION
.
35
3.2.35
S
TABLE
TRANSFORMATION
OF
N
ICOTIANA
TABACUM
PLANTS
.
36
3.2.36
P
LANT
RESISTANCE
ASSAYS
PATHOGEN
INFECTION
.
36
3.2.37
R
ECAPTURE
OF
BACTERIA
FROM
INFECTED
PLANTS
.
37
3.2.38
SUMO
DIGESTION
.
37
3.2.39
T
ESTING
CHLOROPLAST
TRANSFORMATION
CONSTRUCTS
IN
E
.
COLI
.
37
3.2.40
C
ONFOCAL
MICROSCOPY
.
38
3.2.41
T
RANSMISSION
ELECTRON
MICROSCOPY
(TEM).
38
3.3
R
ESULTS
.
39
3.3.1
G
ENERATION
AND
VALIDATION
OF
TRANSPLASTOMIC
PLANTS
THAT
CONSTITUTIVELY
PRODUCE
F
AMP
S
.
39
3.3.2
P
LANT
PHENOTYPES
CAUSED
BY
STRONG
CONSTITUTIVE
EXPRESSION
OF
F
AMP
GENES
.
43
3.3.3
F
AMP
ACCUMULATION
IN
CONSTITUTIVELY
EXPRESSING
PLANTS
.
43
3.3.4
T
HE
INDUCIBLE
RA
MP
ER
SYSTEM
.
44
3.3.5
P
LANT
PHENOTYPES
OF
RA
MP
ER::
F
AMP
PLANTS
.
49
3.3.6
F
AMP
BACKGROUND
LEVELS
IN
RA
MP
ER
AND
P
T
7
PLANTS
.
50
3.3.7
RA
MP
ER
INDUCTION
OF
F
AMP
PRODUCTION
BY
THEOPHYLLINE
.
50
3.3.8
T
HE
EFFECT
OF
THEOPHYLLINE
ON
PLANT
PHENOTYPES
.
51
3.3.9
T
HE
EFFECT
OF
LEAF
AGE
ON
ACCUMULATION
.
51
3.3.10
F
AMP
EXTRACTION
UNDER
NATIVE
CONDITIONS
.
51
3.3.11
T
HE
EFFECT
OF
F
AMP
ACCUMULATION
ON
CHLOROPLAST
INTEGRITY
.
56
3.3.12
E
THANOL
-
INDUCIBLE
F
AMP
PRODUCTION
.
56
3.3.13
F
AMP
EXTRACTION
WITH
DIFFERENT
BUFFERS
AND
DETERGENTS
.
62
3.3.14
D
ENATURING
EXTRACTION
OF
3-N
OVESCANDIN
-5
.
64
3.3.15
T
HE
EFFECT
OF
DETERGENTS
,
DENATURING
AND
ACETIC
ACID
ON
THE
EXTRACTION
OF
3-NOVESCANDIN-5.64
3.3.16
H
IS
-
TAG
PURIFICATION
.
65
3.3.17
I
N
VITRO
ANTIMICROBIAL
ACTIVITY
OF
F
AMP
S
.
65
IV
3.3.18
S
USCEPTIBILITY
OF
F
AMP
PRODUCING
RA
MP
ER
PLANTS
TO
PATHOGENS
.
67
3.3.19
P
ATHOGEN
INDUCIBLE
F
AMP
PRODUCTION
.
72
3.3.20
SUMO
FUSIONS
FOR
ENHANCED
SOLUBILITY
AND
REDUCED
TOXICITY
.
76
3.3.21
A
CCUMULATION
OF
SUMO
FUSIONS
.
77
3.3.22
E
XTRACTION
OF
SUMO-3-W
AMDEPOSIN
-5
.
78
3.3.23
SUMO
REMOVAL
.
78
3.3.24
E
THANOL
-
INDUCIBLE
SUMO-3-W
AMDEPOSIN
-5
.
82
3.3.25
SUMO-HA-WAM1
TESTING
IN
E.
COLI
.
82
3.3.26
C
LEAVAGE
OF
SUMO-HA-WAM1
FROM
E.
COU
.
82
3.4
D
ISCUSSION
.
83
3.4.1
F
AMP
PRODUCTION
IN
CHLOROPLASTS
.
83
3.4.2
T
HE
INDUCIBLE
RA
MP
ER
SYSTEM
FOR
F
AMP
PRODUCTION
.
86
3.4.3
L
EAKY
ACTIVATION
OF
THE
T7
PROMOTER
.
86
3.4.4
RA
MP
ER
INDUCTION
OF
F
AMP
PRODUCTION
.
87
3.4.5
E
T
OH
INDUCIBLE
EXPRESSION
OF
F
AMP
S
.
88
3.4.6
E
XTRACTION
AND
PURIFICATION
OF
F
AMP
S
.
89
3.4.7
I
N
VITRO
ANTIMICROBIAL
ACTIVITY
OF
F
AMP
S
.
90
3.4.8
D
ISEASE
RESISTANCE
OF
F
AMP
PRODUCING
PLANTS
.
90
3.4.9
SUMO
AS
CARRIER
PROTEIN
FOR
(
F
)AMP
PRODUCTION
.
92
3.5
C
ONCLUSION
.
95
4
GENERAL
DISCUSSION
96
R
EFERENCES
.
98
SUPPLEMENTARY
FIGURES
.
105
ALLGEMEINVERSTANDLICHE
ZUSAMMENFASSUNG
IN
DEUTSCHER
SPRACHE
.
109
CURRICULUM
VITAE
.
110
P
UBLICATIONS
.
112
C
ONFERENCE
ATTENDANCE
.
112
S
CHOLARSHIPS
AND
PRIZES
.
112
A
CKNOWLEDGEMENTS
.
113
V |
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| author | Hoelscher, Matthijs Pieter |
| author_GND | (DE-588)1219005088 |
| author_facet | Hoelscher, Matthijs Pieter |
| author_role | aut |
| author_sort | Hoelscher, Matthijs Pieter |
| author_variant | m p h mp mph |
| building | Verbundindex |
| bvnumber | BV047328040 |
| ctrlnum | (OCoLC)1225284437 (DE-599)DNB1221382128 |
| discipline | Biologie |
| discipline_str_mv | Biologie |
| format | Thesis Book |
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| genre_facet | Hochschulschrift |
| id | DE-604.BV047328040 |
| illustrated | Illustrated |
| index_date | 2024-07-03T17:31:44Z |
| indexdate | 2024-07-10T09:09:01Z |
| institution | BVB |
| language | English |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-032730632 |
| oclc_num | 1225284437 |
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| physical | xiii, 114 Seiten Illustrationen 30 cm |
| publishDate | 2020 |
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| spelling | Hoelscher, Matthijs Pieter Verfasser (DE-588)1219005088 aut The production of antimicrobial polypeptides in chloroplasts Matthijs Pieter Hoelscher Potsdam August 2020 xiii, 114 Seiten Illustrationen 30 cm txt rdacontent n rdamedia nc rdacarrier Dissertation Universität Potsdam 2020 (DE-588)4113937-9 Hochschulschrift gnd-content B:DE-101 application/pdf https://d-nb.info/1221382128/04 Inhaltsverzeichnis DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=032730632&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Hoelscher, Matthijs Pieter The production of antimicrobial polypeptides in chloroplasts |
| subject_GND | (DE-588)4113937-9 |
| title | The production of antimicrobial polypeptides in chloroplasts |
| title_auth | The production of antimicrobial polypeptides in chloroplasts |
| title_exact_search | The production of antimicrobial polypeptides in chloroplasts |
| title_exact_search_txtP | The production of antimicrobial polypeptides in chloroplasts |
| title_full | The production of antimicrobial polypeptides in chloroplasts Matthijs Pieter Hoelscher |
| title_fullStr | The production of antimicrobial polypeptides in chloroplasts Matthijs Pieter Hoelscher |
| title_full_unstemmed | The production of antimicrobial polypeptides in chloroplasts Matthijs Pieter Hoelscher |
| title_short | The production of antimicrobial polypeptides in chloroplasts |
| title_sort | the production of antimicrobial polypeptides in chloroplasts |
| topic_facet | Hochschulschrift |
| url | https://d-nb.info/1221382128/04 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=032730632&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT hoelschermatthijspieter theproductionofantimicrobialpolypeptidesinchloroplasts |
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