Molecular cloning and characterization of a regulatory subunit of a cAMP dependent protein kinase from the filarial parasite Onchocerca volvulus (Leuckart, 1893):
Saved in:
| Main Author: | |
|---|---|
| Format: | Thesis Book |
| Language: | English |
| Published: |
2001
|
| Subjects: | |
| Online Access: | Inhaltsverzeichnis |
| Physical Description: | IX, 107 S. Ill., graph. Darst. : 21 cm |
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MARC
| LEADER | 00000nam a2200000 c 4500 | ||
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| 001 | BV014319652 | ||
| 003 | DE-604 | ||
| 005 | 20021113 | ||
| 007 | t | ||
| 008 | 020528s2001 gw ad|| m||| 00||| eng d | ||
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| 040 | |a DE-604 |b ger |e rakddb | ||
| 041 | 0 | |a eng | |
| 044 | |a gw |c DE | ||
| 049 | |a DE-355 |a DE-12 |a DE-83 |a DE-188 | ||
| 100 | 1 | |a Djoha, Sylvain |d 1969- |e Verfasser |0 (DE-588)123497809 |4 aut | |
| 245 | 1 | 0 | |a Molecular cloning and characterization of a regulatory subunit of a cAMP dependent protein kinase from the filarial parasite Onchocerca volvulus (Leuckart, 1893) |c by Sylvain Djoha |
| 264 | 1 | |c 2001 | |
| 300 | |a IX, 107 S. |b Ill., graph. Darst. : 21 cm | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
| 338 | |b nc |2 rdacarrier | ||
| 502 | |a Hamburg, Univ., Diss., 2001 | ||
| 650 | 0 | 7 | |a Proteinkinase C |0 (DE-588)4306407-3 |2 gnd |9 rswk-swf |
| 650 | 0 | 7 | |a Cyclo-AMP |0 (DE-588)4148493-9 |2 gnd |9 rswk-swf |
| 650 | 0 | 7 | |a Onchocerca volvulus |0 (DE-588)4210380-0 |2 gnd |9 rswk-swf |
| 650 | 0 | 7 | |a Molekularbiologie |0 (DE-588)4039983-7 |2 gnd |9 rswk-swf |
| 655 | 7 | |0 (DE-588)4113937-9 |a Hochschulschrift |2 gnd-content | |
| 689 | 0 | 0 | |a Onchocerca volvulus |0 (DE-588)4210380-0 |D s |
| 689 | 0 | 1 | |a Proteinkinase C |0 (DE-588)4306407-3 |D s |
| 689 | 0 | 2 | |a Cyclo-AMP |0 (DE-588)4148493-9 |D s |
| 689 | 0 | 3 | |a Molekularbiologie |0 (DE-588)4039983-7 |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=009824116&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |3 Inhaltsverzeichnis |
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Record in the Search Index
| _version_ | 1807322656187351040 |
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| adam_text |
TABLE
OF
CONTENTS
LIST
OF
ABBREVIATIONS
.VIII
1
INTRODUCTION
.
1
1.1
NEMATODES
.
1
1.2
FILARIASIS
.
1
1.3
ONCHOCERCIASIS
.
2
1.3.1
THE
DISEASE
AND
ITS
PREVALENCE
.
2
1.3.2
O.
VOLVULUS
PARASITOLOGY
AND
LIFE
CYCLE
.
3
1.3.3
DIAGNOSIS
OF
ONCHOCERCIASIS
.
8
1.3.4
TREATMENT
OF
ONCHOCERCIASIS
.
9
1.3.5
CONTROL
OF
ONCHOCERCIASIS
.
10
1.3.6
ONCHOCERCIASIS
IN
CAMEROON.
12
1.3.7
NEW
TOOLS
FOR
THE
FIGHT
AGAINST
ONCHOCERCIASIS
.
15
1.4
PROTEIN
KINASES
.
15
1.4.1
TYPES
OF
PROTEIN
KINASES
.
15
1.4.2
G-PROTEINS
AND
C
AMP-SIGNALING
PATHWAY
.
16
1.4.3
THE
CAMP-DEPENDENT
PROTEIN
KINASE
.
19
1.5
OBJECTIVES
.
24
2
MATERIALS
.
26
2.1
LABORATORY
INSTRUMENTS
.
26
2.2
CHEMICALS
AND
BIOLOGICAL
REAGENTS
.
26
2.2.1
IMMUNOLOGICAL
REAGENTS
.
27
2.2.2
ELECTROPHORESIS
AND
WESTERN
BLOT
REAGENTS
.
27
2.2.3
ENZYMES
USED
TO
MODIFY
DNA
AND
RNA
FRAGMENTS
.
28
2.2.4
KITS
.
28
2.2.5
VECTORS,
BACTERIA
AND
CELLS
USED
.
29
2.3
PARASITE
MATERIALS
.
29
3
METHODS
.
31
3.1
SCREENING
OF
IGTL
1
LIBRARIES
.
31
3.1.1
PREPARATION
OF
BACTERIA
FOR
PLATING
.
31
3.1.2
IMMUNOSCREENING
OF
XGTL
1
CDNA
LIBRARIES
OF
O.
VOLVULUS
.
31
3.2
SEQUENCE
ANALYSIS
AND
IDENTIFICATION
OF
CLONES
.
33
3.2.1
SYNTHESIS
OF
OLIGONUCLEOTIDES
.
33
3.2.2
POLYMERASE
CHAIN
REACTION
(PCR)
.
33
3.2.3
SEQUENCING
OF
SELECTED
CLONES
AND
ANALYSIS
.
34
3.2.4
SUB-CLONING
OF
PCR
FRAGMENTS
IN
THE
TOPO
PCR
AND
PCR
2.1
VECTOR
.
34
3.3
TRANSFORMATION
PROCEDURE
.
35
3.3.1
PREPARATION
OF
COMPETENT
E.
COLI
.
35
3.3.2
TRANSFORMATION
OF
COMPETENT
CELLS
.
35
3.4
DNA
PREPARATION
.
36
3.5
ELECTROPHORESIS
OF
DNA
ON
AGAROSE
GEL
.
37
3.6
PHENOL-CHLOROFORM
EXTRACTION
AND
ETHANOL
PRECIPITATION
OF
DNA.37
3.7
SCREENING
OF
THE
XZAPII
CDNA
LIBRARY
.
38
3.8
IN
VIVO
EXCISION
OF
PLASMID
VECTORS
FROM
BACTERIOPHAGE
XZAPII
ARMS
.
39
3.9
RESTRICTION
DIGESTION
OF
RECOMBINANT
PBLUESCRIPT
PHAGEMIDS
.
40
3.10
EXPRESSION
OF
THE
RECOMBINANT
PROTEIN
.
40
3.10.1
PREPARATION
OF
CDNA
INSERTS
.
40
3.10.2 PREPARATION
OF
THE
P
JC45
VECTOR
FOR
EXPRESSION
PURPOSES
.
41
3.10.3
TRANSFORMATION
REACTION
WITH
THE
RECOMBINANT
PJC45
VECTOR
.
41
3.10.4 INDUCTION
TESTS
.
42
3.11
LARGE
SCALE
EXPRESSION
OF
THE
RECOMBINANT
OV
RPKA
.
43
3.11.1
PREPARATION
OF
CELL
EXTRACT
.
43
3.11.2
PROTEIN
GEL
ELECTROPHORESIS
.
43
3.12
PROTEIN
STAINING
IN
GELS
.
44
3.13
WESTERN
BLOTTING
.
45
3.14
IMMUNODETECTION
.
46
3.15
PURIFICATION
OF
RECOMBINANT
OV
RPKA
.
47
3.16
PRODUCTION
OF
OV
RPKA
SPECIFIC
ANTISERUM
.
48
3.17
ANTIGEN
PREPARATION
FROM
ADULT
O.
VOLVULUS
WORMS
.
49
3.18
IMMUNOLOCALIZATION
OF
OV
RPKA
IN
PARASITE
TISSUES
.
49
4
RESULTS
.
51
4.1
SCREENING
OF
O.
VOLVULUS
IGTL
1
CDNA
LIBRARIES
.
51
4.1.1
ISOLATION
OF
A
CDNA
FRAGMENT
CODING
FOR
THE
OV
RPKA
.
51
4.1.2
GENERATION
OF
THE
CDNA
PROBE
.
52
4.2 SCREENING
OF
THE
KZAP
CDNA
LIBRARY
.
53
4.3
THE
COMPLETE
CDNA
SEQUENCE
OF
THE
OV
RPKA
.
55
4.3.1
FEATURES
OF
THE
1435
BP
CDNA
.
55
4.3.2
A
REGULATORY
SUBUNIT
OF
A
CAMP-DEPENDENT
PROTEIN
KINASE
OF
O.
VOLVULUS
.
57
4.4.
RECOMBINANT
EXPRESSION
AND
PURIFICATION
OF
THE
OV
RPKA
.
62
4.4.1
RECOMBINANT
EXPRESSION
OF
THE
OV
RPKA
.
62
4.4.2
PURIFICATION
OF
THE
RECOMBINANT
OV
RPKA
.
63
4.5
IDENTIFICATION
OF
THE
NATIVE
OV
RPKA
.
65
4.6
REACTIVITY
OF
OV
RPKA
WITH
SERA
FROM
PATIENTS
WITH
ONCHOCERCIASIS
AND
OTHER
FILARIAL
INFECTIONS
.
67
4.7
IMMUNOLOCALIZATION
OF
THE
NATIVE
OV
RPKA
IN
O.
VOLVULUS
TISSUES
.
71
4.7.1
IMMUNOHISTOCHEMISTRY
.
71
4.7.2
MICROFILARIAE
AND
EMBRYOS
.
72
4.7.3
ADULT
O.
VOLVULUS
FILARIAE
.
75
4.7.4
OTHER
FILARIAE
.
81
5
DISCUSSION
.
83
5.1
ISOLATION
OF
THE
1435
CDNA
AND
EXPRESSION
OF
THE
RECOMBINANT
PROTEIN
.
83
5.2
STRUCTURAL
DOMAINS
AND
PREDICTED
FEATURES
OF
OV
RPKA
.
83
5.3
RELATIONSHIPS
BETWEEN
LOCALIZATION
AND
PREDICTED
FUNCTIONS
OF
OVRPKA
.
90
5.4
SERODIAGNOSTIC
ASSESSMENT
.
93
5.5
CONCLUSIONS
.
94
6
SUMMARY.
96
7
REFERENCES
.
98
ACKNOWLEDGEMENTS
CURRICULUM
VITAE
LINGUISTIC
STATEMENT
ERKLARUNG |
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| author_GND | (DE-588)123497809 |
| author_facet | Djoha, Sylvain 1969- |
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| genre_facet | Hochschulschrift |
| id | DE-604.BV014319652 |
| illustrated | Illustrated |
| indexdate | 2024-08-14T00:59:56Z |
| institution | BVB |
| language | English |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-009824116 |
| oclc_num | 51521428 |
| open_access_boolean | |
| owner | DE-355 DE-BY-UBR DE-12 DE-83 DE-188 |
| owner_facet | DE-355 DE-BY-UBR DE-12 DE-83 DE-188 |
| physical | IX, 107 S. Ill., graph. Darst. : 21 cm |
| publishDate | 2001 |
| publishDateSearch | 2001 |
| publishDateSort | 2001 |
| record_format | marc |
| spelling | Djoha, Sylvain 1969- Verfasser (DE-588)123497809 aut Molecular cloning and characterization of a regulatory subunit of a cAMP dependent protein kinase from the filarial parasite Onchocerca volvulus (Leuckart, 1893) by Sylvain Djoha 2001 IX, 107 S. Ill., graph. Darst. : 21 cm txt rdacontent n rdamedia nc rdacarrier Hamburg, Univ., Diss., 2001 Proteinkinase C (DE-588)4306407-3 gnd rswk-swf Cyclo-AMP (DE-588)4148493-9 gnd rswk-swf Onchocerca volvulus (DE-588)4210380-0 gnd rswk-swf Molekularbiologie (DE-588)4039983-7 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Onchocerca volvulus (DE-588)4210380-0 s Proteinkinase C (DE-588)4306407-3 s Cyclo-AMP (DE-588)4148493-9 s Molekularbiologie (DE-588)4039983-7 s DE-604 DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=009824116&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Djoha, Sylvain 1969- Molecular cloning and characterization of a regulatory subunit of a cAMP dependent protein kinase from the filarial parasite Onchocerca volvulus (Leuckart, 1893) Proteinkinase C (DE-588)4306407-3 gnd Cyclo-AMP (DE-588)4148493-9 gnd Onchocerca volvulus (DE-588)4210380-0 gnd Molekularbiologie (DE-588)4039983-7 gnd |
| subject_GND | (DE-588)4306407-3 (DE-588)4148493-9 (DE-588)4210380-0 (DE-588)4039983-7 (DE-588)4113937-9 |
| title | Molecular cloning and characterization of a regulatory subunit of a cAMP dependent protein kinase from the filarial parasite Onchocerca volvulus (Leuckart, 1893) |
| title_auth | Molecular cloning and characterization of a regulatory subunit of a cAMP dependent protein kinase from the filarial parasite Onchocerca volvulus (Leuckart, 1893) |
| title_exact_search | Molecular cloning and characterization of a regulatory subunit of a cAMP dependent protein kinase from the filarial parasite Onchocerca volvulus (Leuckart, 1893) |
| title_full | Molecular cloning and characterization of a regulatory subunit of a cAMP dependent protein kinase from the filarial parasite Onchocerca volvulus (Leuckart, 1893) by Sylvain Djoha |
| title_fullStr | Molecular cloning and characterization of a regulatory subunit of a cAMP dependent protein kinase from the filarial parasite Onchocerca volvulus (Leuckart, 1893) by Sylvain Djoha |
| title_full_unstemmed | Molecular cloning and characterization of a regulatory subunit of a cAMP dependent protein kinase from the filarial parasite Onchocerca volvulus (Leuckart, 1893) by Sylvain Djoha |
| title_short | Molecular cloning and characterization of a regulatory subunit of a cAMP dependent protein kinase from the filarial parasite Onchocerca volvulus (Leuckart, 1893) |
| title_sort | molecular cloning and characterization of a regulatory subunit of a camp dependent protein kinase from the filarial parasite onchocerca volvulus leuckart 1893 |
| topic | Proteinkinase C (DE-588)4306407-3 gnd Cyclo-AMP (DE-588)4148493-9 gnd Onchocerca volvulus (DE-588)4210380-0 gnd Molekularbiologie (DE-588)4039983-7 gnd |
| topic_facet | Proteinkinase C Cyclo-AMP Onchocerca volvulus Molekularbiologie Hochschulschrift |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=009824116&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT djohasylvain molecularcloningandcharacterizationofaregulatorysubunitofacampdependentproteinkinasefromthefilarialparasiteonchocercavolvulusleuckart1893 |