Protein semisynthesis and engineering by expressed protein ligation:
Gespeichert in:
| 1. Verfasser: | |
|---|---|
| Format: | Buch |
| Sprache: | English |
| Veröffentlicht: |
2002
|
| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis |
| Beschreibung: | Leipzig, Univ., Diss., 2002 |
| Beschreibung: | 158 S. Ill., graph. Darst. |
Internformat
MARC
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| 100 | 1 | |a Machova, Zuzana |e Verfasser |4 aut | |
| 245 | 1 | 0 | |a Protein semisynthesis and engineering by expressed protein ligation |c vorgelegt von Zuzana Machova |
| 264 | 1 | |c 2002 | |
| 300 | |a 158 S. |b Ill., graph. Darst. | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
| 338 | |b nc |2 rdacarrier | ||
| 500 | |a Leipzig, Univ., Diss., 2002 | ||
| 650 | 0 | 7 | |a Proteindesign |0 (DE-588)4346801-9 |2 gnd |9 rswk-swf |
| 650 | 0 | 7 | |a Ligation |0 (DE-588)4723826-4 |2 gnd |9 rswk-swf |
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| 689 | 0 | 0 | |a Proteindesign |0 (DE-588)4346801-9 |D s |
| 689 | 0 | 1 | |a Ligation |0 (DE-588)4723826-4 |D s |
| 689 | 0 | |5 DE-604 | |
| 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=010212402&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |3 Inhaltsverzeichnis |
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Datensatz im Suchindex
| _version_ | 1807864751880929280 |
|---|---|
| adam_text |
TABLE
OF
CONTENTS
1
TABLE
OF
CONTENTS
TABLE
OF
CONTENTS
1
ABBREVIATIONS
5
BIBLIOGRAPHISCHE
BESCHREIBUNG
9
SUMMARY
11
ZUSAMMENFASSUNG
17
CHAPTER
1
23
EXPRESSED
PROTEIN
LIGATION
FOR
PROTEIN
SEMISYNTHESIS
AND
ENGINEERING
23
1.1
ABSTRACT
24
1.2
INTRODUCTION
25
1.3
EXPRESSED
PROTEIN
LIGATION
26
1.3.1
INTEINS
AND
PROTEIN
SPLICING
27
1.3.2
IMPACT
PURIFICATION
SYSTEM 29
1.3.3
CHEMICAL
LIGATION
34
1.4
PROTEIN
ENGINEERING
BY
EXPRESSED
PROTEIN
LIGATION
36
1.4.1
SITE-SPECIFIC
PROTEIN
MODIFICATIONS
36
1.4.2
BACKBONE
CYCLIZATION
AND
POLYMERIZATION
OF
PROTEINS
39
1.4.3
SEGMENTAL
ISOTOPIC
LABELLING
41
1.4.4
PRODUCTION
OF
CYTOTOXIC
PROTEINS
43
1.4.5
STUDIES
OF
PROTEIN-PROTEIN
INTERACTIONS 44
1.4.6
APPLICATION
OF
GREEN
FLUORESCENT
PROTEIN
IN
EPL
45
1.4.7
EXPRESSED
ENZYMATIC
LIGATION
47
1.5
CONCLUSIONS
48
1.6
REFERENCES
49
CHAPTER
2
57
CELLULAR
INTERNALIZATION
OF
ENHANCED
GREEN
FLUORESCENT
PROTEIN
LIGATED
TO
A
HUMAN
CALCITONIN-BASED
CARRIER
PEPTIDE
57
2.1
ABSTRACT
58
2
TABLE
OF
CONTENTS
2.2
INTRODUCTION
59
2.3
RESULTS
AND
DISCUSSION
61
2.3.1
STRATEGY
61
2.3.2
CLONING,
EXPRESSION,
AND
ISOLATION
OF
EGFP-INTEIN-CBD
FUSION
PROTEIN
61
2.3.3
ISOLATION
OF
THE
EGFP
THIOESTER
63
2.3.4
LIGATION
REACTION
OF
EGFP
THIOESTER
WITH
[C
E
]HCT8-32
65
2.3.5
ELECTROPHORETIC
ANALYSIS
66
2.3.6
CELLULAR
INTERNALIZATION
IN
NASAL
MUCOSA
AS
MONITORED
BY
CLSM
67
2.4 EXPERIMENTAL
SECTION
70
2.4.1
DNA
CONSTRUCTS
70
2.4.2
FUSION
PROTEIN
EXPRESSION
IN
E.
COLL
71
2.4.3
PURIFICATION
AND
ISOLATION
OF
EGFP
THIOESTER
71
2.4.4
SYNTHESIS
OF
HCT-DERIVED
CARRIER
PEPTIDES
72
2.4.5
LIGATION
OF
THE
EGFP
THIOESTER
WITH
[C
8
]HCT8-32
72
2.4.6
INTERNALIZATION
STUDIES
ON
BOVINE
NASAL
MUCOSA
72
2.4.7
VISUALIZATION
BY
CONFOCAL
LASER
SCANNING
MICROSCOPY
73
2.5
ACKNOWLEDGEMENT
73
2.6
REFERENCES
74
CHAPTER
3
79
SEMISYNTHESIS
OF
CARBOXYFLUORESCEIN-LABELLED,
BIOLOGICALLY
ACTIVE
HUMAN
LNTERLEUKIN-8
BY
EXPRESSED
PROTEIN
LIGATION
79
3.1
SUMMARY
80
3.2
INTRODUCTION
81
3.3
EXPERIMENTAL
PROCEDURES
83
3.3.1
DNA
CONSTRUCTS
83
3.3.2
FUSION
PROTEIN
EXPRESSION
IN
E.
COLI
84
3.3.3
PURIFICATION
AND
ISOLATION
OF
HLL-8(1
-54)
THIOESTER
84
3.3.4
SYNTHESIS
OF
HLL-8(55-77)
AND
[K
69
(CF)]HLL-8(55-77)
85
3.3.5
LIGATION
OF
HLL-8(1-77)
AND
[K
69
(CF)]HLL-8(1
-77)
86
3.3.6
CELL
CULTURE
86
3.3.7
CAMP
ENZYME-IMMUNOASSAY
86
TABLE
OF
CONTENTS
3
3.3.8
COMPETITION
BINDING
STUDIES
87
3.3.9
INTERNALIZATION
ASSAY
87
3.4
RESULTS
88
3.4.1
STRATEGY
88
3.4.2
PURIFICATION
OF
HLL-8(1-54)-INTEIN-CBD
FUSION
PROTEIN
88
3.4.3
ISOLATION
OF
THE
HLL-8(1-54)
THIOESTER
91
3.4.4
GENERATION
OF
HLL-8(1-77)
AND
[K
69
(CF)]HLL-8(1
-77)
BY
LIGATION
93
3.4.5
DETERMINATION
OF
THE
BIOLOGICAL
ACTIVITY
OF
[K
69
(CF)]HLL-8(1
-77)
94
3.5
DISCUSSION
96
3.6
ACKNOWLEDGEMENT
99
3.7
REFERENCES
100
CHAPTER
4
107
SEMISYNTHESIS
AND
CHARACTERIZATION
OF
THE
FIRST
ANALOGUES
OF
PRO
NEUROPEPTIDE
Y
107
4.1
ABSTRACT
108
4.2
INTRODUCTION
109
4.3
RESULTS
112
4.3.1
STRATEGY
112
4.3.2
PURIFICATION
OF
FUSION
PROTEINS
112
4.3.3
GENERATION
OF
PROTEIN
THIOESTER
114
4.3.4
STABILITY
OF
PROTEIN
THIOESTER
116
4.3.5
SOLID-PHASE
SYNTHESIS
OF
C-TERMINAL
PRONPY SEGMENTS
116
4.3.6
GENERATION
OF
PRONPY
ANALOGUES
BY
CHEMICAL
LIGATION
119
4.3.7
ANALYSIS
OF
LIGATING
FRAGMENTS
AND
LIGATION
PRODUCTS
119
4.4
DISCUSSION
121
4.5
EXPERIMENTAL
SECTION
124
4.5.1
MATERIALS
124
4.5.2
DNA
CONSTRUCTS
125
4.5.3
FUSION
PROTEIN
EXPRESSION
IN
E.
COLI
125
4.5.4
PURIFICATION
AND
ISOLATION
OF
PROTEIN
THIOESTER
126
4.5.5
ANALYSIS
OF
THE
CLEAVAGE
OF
INTEIN-FUSION
PROTEIN
127
4
TABLE
OF
CONTENTS
4.5.6
THIOESTER
STABILITY
ASSAY
127
4.5.7
SYNTHESIS
OF
PRONPY-DERIVED
PEPTIDES
127
4.5.8
LIGATION
OF
PRONPY-DERIVED
ANALOGUES
129
4.5.9
ANALYSIS
OF
FRAGMENTS
AND
PRODUCTS
BY
WESTERN
AND
DOTBLOTTING
129
4.6
ACKNOWLEDGEMENT
130
4.7
REFERENCES
131
CHAPTER
5
135
EXPRESSED
ENZYMATIC
LIGATION
FOR
THE
SEMISYNTHESIS
OF
CHEMICALLY
MODIFIED
PROTEINS
135
5.1
ABSTRACT
136
5.2
INTRODUCTION
137
5.3
RESULTS
AND
DISCUSSION
139
5.3.1
STRATEGY
139
5.3.2
ISOLATION
OF
THE
PRONPY(1-40)
THIOESTERS
140
5.3.3
V8
PROTEASE-MEDIATED
ENZYMATIC
LIGATION
143
5.3.4
CONCLUSIONS
146
5.4
EXPERIMENTAL
SECTION
147
5.4.1
DNA
CONSTRUCT
147
5.4.2
FUSION
PROTEIN
EXPRESSION
IN
E.
COLL
147
5.4.3
PURIFICATION
AND
ISOLATION
OF
PRONPY(1-40)
THIOESTER
148
5.4.4
SYNTHESIS
OF
PRONPY-DERIVED
PEPTIDES
148
5.4.5
V8
PROTEASE-MEDIATED
ENZYMATIC
LIGATIONS
149
5.5
ACKNOWLEDGEMENT
149
5.6
REFERENCES
150
PUBLICATIONS
153
POSTERS
154
ORAL
PRESENTATIONS
155
CURRICULUM
VITAE
156 |
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| author | Machova, Zuzana |
| author_facet | Machova, Zuzana |
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| author_sort | Machova, Zuzana |
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| bvnumber | BV016526126 |
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| illustrated | Illustrated |
| indexdate | 2024-08-20T00:36:20Z |
| institution | BVB |
| language | English |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-010212402 |
| oclc_num | 633226135 |
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| owner | DE-91 DE-BY-TUM DE-355 DE-BY-UBR |
| owner_facet | DE-91 DE-BY-TUM DE-355 DE-BY-UBR |
| physical | 158 S. Ill., graph. Darst. |
| publishDate | 2002 |
| publishDateSearch | 2002 |
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| record_format | marc |
| spelling | Machova, Zuzana Verfasser aut Protein semisynthesis and engineering by expressed protein ligation vorgelegt von Zuzana Machova 2002 158 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Leipzig, Univ., Diss., 2002 Proteindesign (DE-588)4346801-9 gnd rswk-swf Ligation (DE-588)4723826-4 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Proteindesign (DE-588)4346801-9 s Ligation (DE-588)4723826-4 s DE-604 DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=010212402&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Machova, Zuzana Protein semisynthesis and engineering by expressed protein ligation Proteindesign (DE-588)4346801-9 gnd Ligation (DE-588)4723826-4 gnd |
| subject_GND | (DE-588)4346801-9 (DE-588)4723826-4 (DE-588)4113937-9 |
| title | Protein semisynthesis and engineering by expressed protein ligation |
| title_auth | Protein semisynthesis and engineering by expressed protein ligation |
| title_exact_search | Protein semisynthesis and engineering by expressed protein ligation |
| title_full | Protein semisynthesis and engineering by expressed protein ligation vorgelegt von Zuzana Machova |
| title_fullStr | Protein semisynthesis and engineering by expressed protein ligation vorgelegt von Zuzana Machova |
| title_full_unstemmed | Protein semisynthesis and engineering by expressed protein ligation vorgelegt von Zuzana Machova |
| title_short | Protein semisynthesis and engineering by expressed protein ligation |
| title_sort | protein semisynthesis and engineering by expressed protein ligation |
| topic | Proteindesign (DE-588)4346801-9 gnd Ligation (DE-588)4723826-4 gnd |
| topic_facet | Proteindesign Ligation Hochschulschrift |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=010212402&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
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