Cloning of genes from thermophilic bacteria and their expression in Bacillus subtilis and plants:
Gespeichert in:
| 1. Verfasser: | |
|---|---|
| Format: | Buch |
| Sprache: | German |
| Veröffentlicht: |
1995
|
| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis |
| Beschreibung: | Halle, Univ., Diss., 1995 |
| Beschreibung: | VII, 98 S. Ill., graph. Darst. |
Internformat
MARC
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| 035 | |a (DE-599)BVBBV010698113 | ||
| 040 | |a DE-604 |b ger |e rakddb | ||
| 041 | 0 | |a ger | |
| 044 | |a gw |c DE | ||
| 049 | |a DE-91 |a DE-355 | ||
| 100 | 1 | |a Abd- el Azim, Abd-ElAzim Farouk |e Verfasser |4 aut | |
| 245 | 1 | 0 | |a Cloning of genes from thermophilic bacteria and their expression in Bacillus subtilis and plants |c von Abd-ElAzim Farouk Abd-ElAzim |
| 264 | 1 | |c 1995 | |
| 300 | |a VII, 98 S. |b Ill., graph. Darst. | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
| 338 | |b nc |2 rdacarrier | ||
| 500 | |a Halle, Univ., Diss., 1995 | ||
| 650 | 0 | 7 | |a Proteasen |0 (DE-588)4047521-9 |2 gnd |9 rswk-swf |
| 650 | 0 | 7 | |a Analyse |0 (DE-588)4122795-5 |2 gnd |9 rswk-swf |
| 650 | 0 | 7 | |a Thermoactinomyces vulgaris |0 (DE-588)4402189-6 |2 gnd |9 rswk-swf |
| 650 | 0 | 7 | |a Heubacillus |0 (DE-588)4242465-3 |2 gnd |9 rswk-swf |
| 650 | 0 | 7 | |a Heterologe Genexpression |0 (DE-588)4342015-1 |2 gnd |9 rswk-swf |
| 650 | 0 | 7 | |a Xylose |0 (DE-588)4190397-3 |2 gnd |9 rswk-swf |
| 650 | 0 | 7 | |a Genklonierung |0 (DE-588)4123275-6 |2 gnd |9 rswk-swf |
| 655 | 7 | |0 (DE-588)4113937-9 |a Hochschulschrift |2 gnd-content | |
| 689 | 0 | 0 | |a Thermoactinomyces vulgaris |0 (DE-588)4402189-6 |D s |
| 689 | 0 | 1 | |a Proteasen |0 (DE-588)4047521-9 |D s |
| 689 | 0 | 2 | |a Genklonierung |0 (DE-588)4123275-6 |D s |
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| 943 | 1 | |a oai:aleph.bib-bvb.de:BVB01-007140455 | |
Datensatz im Suchindex
| _version_ | 1807865556912570368 |
|---|---|
| adam_text |
CONTENTS
1.
INTRODUCTION
1
1.1.
PRESENT
USE
OF
BACILLUS
SUBTLLLS
FOR
SECRETION
OF
HETEROLOGOUS
PROTEINS
.
1
1.1.1.
TRANSFORMATION
AND
VECTORS
.
1
1.1.2.
CONSTITUTIVE
VERSUS
INDUCIBLE
PROMOTERS
.2
1.1.3.
SECRETORY
EXPRESSION
SYSTEMS
.2
1.1.4.
THE
PROTEASES
AND
PROTEOLYSIS
.3
1.2.
THAMOACTLNOMYCES
VULGARIS
K94;
A
SOURCE
OF
THERMOSTABLE
ENZYMES
.
4
1.2.1.
THEGENUS
THERMOACTINOMYCES
.4
1.2.2.
ALPHA-AMYLASES
AND
OTHER
EXTRACELLULAR
ENZYMES
.
4
1.3.
THE
A-AMYLASES
AND
CELLULASES
IN
PLANT
PHYSIOLOGY
.
5
1.3.1
STARCH
AND
STARCH
DEGRADATION
BY
AMYLOLYTIC
ENZYMES
IN
CHLOROPLASTS
AND
SEEDS
.
5
1.3.2.
CELLULOSE
AND
CELLULASES
IN
PLANTS
.6
1.4.
THE
BASIC
PREPOSITIONS
TO
PRODUCE
BACTERIAL
PROTEINS
IN
PLANTS
.
7
1.4.1.
PLANT
EXPRESSION
CASSETTES
.
7
1.4.2.
THE
TI
PLASMID
AND
PLANT
TRANSFORMATION
.8
1.4.3.
TARGETING
OF
FOREIGN
PROTEIN
IN
PLANTS
.
8
1.4.4.
PLANT
GENETIC
ENGINEERING;
THE
USE
OF
BACTERIAL
GENES
.9
1.4.5.
BIOFARMING
IN
PLANTS
.
10
2.
MATERIALS
AND
METHODS
.
11
2.1.
SUBSTRATES
AND
REAGENTS
.
11
2.2.
STRAINS
AND
VECTORS
.
11
2.2.1.
STRAINS
.
11
2.2.2.
VECTORS
.
11
2.3.
MEDIA
AND
CULTURE
CONDITIONS
.
12
2.3.1.
MEDIA
.
12
2.3.2.
CULTURE
CONDITIONS
.13
2.4.
DNA
TECHNIQUES
.
13
2.4.1.
DNA
TECHNIQUES
FOR
BACTERIA
.
13
2.4.1.1.
PLASMID
DNA
ISOLATION
.
13
2.4.1.2.
PLASMID
DNA
PURIFICATION.
14
2.4.1.3.
CHROMOSOMAL
DNA
MINI-PREPREPARATION
.
14
2.4.1.4.
DNA
MINIPREPARATION
FROM
BACTERIOPHAGES
.
14
2.4.1.5.
DNA
RESTRICTION
DIGESTION
.
15
ILL
24.1.6.
DNA
EXTRACTION
AND
PURIFICATION
FROM
AGAROSE
GEL
.15
2.4.1.7.
FILLING
OF
DNA
FRAGMENT
ENDS
.15
2.4.1.8.
DEPHOSPHORYLATION
OF
LINEARIZED
PLASMID
DNA
.16
2.4.1.9.
DNA
LIGATION
.16
24.1.10.
DNA
SEQUENCING
.16
2.4.1.11.
PCR
.16
2
4.1.12.
THE
TRANSFORMATION
OF
DNA
INTO
E.
COLI
.16
2.4.1.13.
THE
TRANSFORMATION
OF
DNA
INTO
B.
SUBTILIS
.16
2
4.1.14.
SOUTHERN
BLOT
.16
2.4.2.
DNA
TECHNIQUES
FOR
PLANTS
.
17
2
4.2.1.
AGROBACTERIUM
TUMEFACIENS
DNA
TRANSFER
.
17
2
4.2.2.
TOTAL
DNA
FROM
AGROBACTERIUM
TUMEFACEINS
.17
2.4.2.3.
GENOMIC
DNA
ISOLATION
FROM
TOBACCO
PLANTS
.
17
2.5.
RNA
TECHNIQUES
.18
2.5.1.
RNA
TECHNIQUES
FOR
BACERIA
.18
2.5.1.1.
RNA
ISOLATION
.18
2.5.2.
RNA
TECHNIQUES
FOR
PLANTS
.
19
2.5.2.1.
RNA
FROM
PLANT
LEAVES
.
19
2.5.2.2.
RNA
FROM
TOBACCO
SEEDS
.
19
2.5.2.3.
RNA
ELECTROPHORESIS
.
19
2.5.2.4.
RNA
TRANSFER
(NORTHERN-BLOT)
.
20
2.5.2.5.
RNA
BLOT
HYBRIDIZATION
.
20
2.6.
PLANT
TRANSFORMATION
.
20
2.7.
ASSAY
AND
CHARACTERIZATION
OF
PROTEINS
.
20
2
7.1.
PROTEIN
EXTRACTIONS
.
20
2.7.1.1.
ISOLATION
OF
TOBACCO
CHLOROPLASTS
.
20
2.7.1.2.
ISOLATION
OF
PROTEINS
FROM
THE
INTERCELLULAR
FLUID
.
21
2.7.1.3.
PROTEIN
EXTRACTION
FROM
PLANT
LEAVES
AND
SEEDS
.
21
2.7.2.
PROTEIN
DETERMINATION
.
21
2.7.3.
PROTEIN
CHARACTERIZATION
.
21
2.7.3.1
.
SDS-POLYACRYLAMIDE
GEL
ELECTROPHORESIS
.
21
2.7.3.2.
ELECTROPHORETIC
TRANSFER
AND
IMMUNOSTAINING
.
22
2.7.4.
ENZYME
ACTIVITY
ASSAYS
.
22
2.7.4.1.
A-AMYLASE
ACTIVITY
ASSAYS
.
22
2.7.4.2.
PROTEASE
ACTIVITY
ASSAYS
.
23
2.7.4.3.
CELLULASE
ACTIVITY
ASSAYS
.
23
2.7.4.4.
HPLC
ANALYSIS
.
23
IV
3.
RESULTS
.
24
3.1.
CLONING
OF
A
THERMOLYSIN-LIKE
PROTEASE
GENE
.
24
3.1.1.
SEQUENCING
AND
HOMOLOGIES
.
25
3.1.2.
EXPRESSION
IN
B.
SUBTILIS
.
29
3.1.3.
LOCALIZATION
OF
THE
PROTEASE
TIPE
1
(TV)
IN
B.
SUBTILIS
.
30
3.1.4.
PRELIMINARY
ENZYME
CHARACTERIZATION
.
30
3.1.4.1.
PH-OPTIMUM
.
31
3.1.4.2.
T
EMPERATURE
RESPONSES
.32
3.1.4.3.
INHIBITOR
SENSITIVITY
.32
3.2.
APPROACH
TO
CLONE
THE
THERMITASE
GENE
[TTHE1(TV)]
.
33
3.2.1.
PCR
CLONING
.
33
3.2.2.
DNA
SEQUENCING
.
34
3.2.3.
SCREENING
OF
THE
EMBL4
LIBRARY
.
34
3.3.
USE
OF
THE
XYLR/XYLA
REPRESSOR-REGULATORY
SYSTEM
FOR
SECRETORY
EXPRESSION
IN
B.
SUBTILIS
.
35
3.3.1.
CONSTRUCTION
OF
XYL-AMYTV
AND
XYL-AMYBA
EXPRESSION
CASSETTES
.
36
3.3.2.
GENE
INDUCTION
AND
AMYLASE
SECRETION
.
37
3.3.3.
GLUCOSE
REPRESSION
.
38
3.3.4.
RELATIVE
YIELD
.
40
3.3.5.
LOCALIZATION
OF
THE
INDUCED
AMYTV
PROTEIN
.
41
3.4.
EXPRESSION
OF
BACTERIAL
GENES
IN
PLANTS
.
42
3.4.1.
AMYTV
EXPRESSION
AND
TARGETING
INTO
TOBACCO
CHLOROPLASTS
.
42
3.4.1.1.
THE
CAMV35S-FNR-TP
PROMOTER/LEADER
PEPTIDE
CASSETTE
.
42
3.4.1.2.
TRANSFORMATION
AND
TARGETING
INTO
THE
CHLOROPLASTS.
43
3.4.1.3.
ENZYME
ASSAY
.
43
3.4.1.4.
NORTHERN
ANALYSIS
.
43
3.4.1.5.
WESTERN
ANALYSIS
.
44
3.4.2.
EXPRESSION
OF
CELZ
OF
CLOSTRIDIUM
STERCORARIUM
IN
TOBACCO
.
45
3.4.2.1
.
EXPRESSION
VECTOR
BIN-CE/ZF
AND
TRANSFORMATION
.
45
3.4.2.2.
CELZ
EXPRESSION
AND
LOCALIZATION
IN
TOBACCO
LEAVES
.
46
3.4.3.
SEED
SPECIFIC
EXPRESSION;
THE
LEB4
AND
USP
GENE
PROMOTERS
.
49
3.4.3.1.
LEB4-AMYTV;
TRANSFORMATION,
EXPRESSION
ASSAYS
AND
PRODUCT
CHARACTERIZATION
.
49
3.4.3.2.
USP-AMYTV;
TRANSFORMATION
AND
EXPRESSION
ASSAYS
.
55
3.4.3.3.
USP-CE/Z,
TRANSFORMATION
AND
EXPRESSION
ASSAYS
.
57
3.4.3.4.
LEB4
PROMOTER-AMYSA;
TRANSFORMATION
AND
EXPRESSION
ASSAYS
.59
3.4.3.5.
INHERITANCE
OF
CELZ
AND
AMYTV,
IN
USP
AND
LEB4
CONSTRUCTS
.
61
3.4.3.6.
RELATIVE
CELZ
EXPRESSION
IN
APOPLAST
AND
SEEDS
OF
TOBACCO
.
63
_
V
4.
DISCUSSION
.
65
4.1.
THE
PROTEASES
OF
T.
VULGARIS-,
CLONING
INTO
B.
SUBTIHS
.
65
4.2.
INDUCIBLE
AMYTV
EXPRESSION
AND
SECRETION;
IMPLICATION
FOR
THE
USE
OF
THE
XYLA
INDUCIBLE
PROMOTER
CASSETTE
FOR
PROTEIN
SECRETION
IN
B.
SUBTULS
.
68
4.3.
CAMV
35S
PROMOTER
EXPRESSION
OF
AMYTV
AND
CELZ
.
69
4.
4
SEED
SPECIFIC
EXPRESSION
OF
AMYTV
AND
CELZ
.
71
5.
SUMMARY
.
75
5.1.
SUMMARY
.
75
5.2.
ZUSAMMENFASSUNG
.
77
6.
REFERENCES
.
80 |
| any_adam_object | 1 |
| author | Abd- el Azim, Abd-ElAzim Farouk |
| author_facet | Abd- el Azim, Abd-ElAzim Farouk |
| author_role | aut |
| author_sort | Abd- el Azim, Abd-ElAzim Farouk |
| author_variant | e a a e f a eaaef eaaefa |
| building | Verbundindex |
| bvnumber | BV010698113 |
| ctrlnum | (OCoLC)632673737 (DE-599)BVBBV010698113 |
| format | Book |
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| genre | (DE-588)4113937-9 Hochschulschrift gnd-content |
| genre_facet | Hochschulschrift |
| id | DE-604.BV010698113 |
| illustrated | Illustrated |
| indexdate | 2024-08-20T00:49:06Z |
| institution | BVB |
| language | German |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-007140455 |
| oclc_num | 632673737 |
| open_access_boolean | |
| owner | DE-91 DE-BY-TUM DE-355 DE-BY-UBR |
| owner_facet | DE-91 DE-BY-TUM DE-355 DE-BY-UBR |
| physical | VII, 98 S. Ill., graph. Darst. |
| publishDate | 1995 |
| publishDateSearch | 1995 |
| publishDateSort | 1995 |
| record_format | marc |
| spelling | Abd- el Azim, Abd-ElAzim Farouk Verfasser aut Cloning of genes from thermophilic bacteria and their expression in Bacillus subtilis and plants von Abd-ElAzim Farouk Abd-ElAzim 1995 VII, 98 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Halle, Univ., Diss., 1995 Proteasen (DE-588)4047521-9 gnd rswk-swf Analyse (DE-588)4122795-5 gnd rswk-swf Thermoactinomyces vulgaris (DE-588)4402189-6 gnd rswk-swf Heubacillus (DE-588)4242465-3 gnd rswk-swf Heterologe Genexpression (DE-588)4342015-1 gnd rswk-swf Xylose (DE-588)4190397-3 gnd rswk-swf Genklonierung (DE-588)4123275-6 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Thermoactinomyces vulgaris (DE-588)4402189-6 s Proteasen (DE-588)4047521-9 s Genklonierung (DE-588)4123275-6 s DE-604 Xylose (DE-588)4190397-3 s Analyse (DE-588)4122795-5 s Heterologe Genexpression (DE-588)4342015-1 s Heubacillus (DE-588)4242465-3 s DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=007140455&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Abd- el Azim, Abd-ElAzim Farouk Cloning of genes from thermophilic bacteria and their expression in Bacillus subtilis and plants Proteasen (DE-588)4047521-9 gnd Analyse (DE-588)4122795-5 gnd Thermoactinomyces vulgaris (DE-588)4402189-6 gnd Heubacillus (DE-588)4242465-3 gnd Heterologe Genexpression (DE-588)4342015-1 gnd Xylose (DE-588)4190397-3 gnd Genklonierung (DE-588)4123275-6 gnd |
| subject_GND | (DE-588)4047521-9 (DE-588)4122795-5 (DE-588)4402189-6 (DE-588)4242465-3 (DE-588)4342015-1 (DE-588)4190397-3 (DE-588)4123275-6 (DE-588)4113937-9 |
| title | Cloning of genes from thermophilic bacteria and their expression in Bacillus subtilis and plants |
| title_auth | Cloning of genes from thermophilic bacteria and their expression in Bacillus subtilis and plants |
| title_exact_search | Cloning of genes from thermophilic bacteria and their expression in Bacillus subtilis and plants |
| title_full | Cloning of genes from thermophilic bacteria and their expression in Bacillus subtilis and plants von Abd-ElAzim Farouk Abd-ElAzim |
| title_fullStr | Cloning of genes from thermophilic bacteria and their expression in Bacillus subtilis and plants von Abd-ElAzim Farouk Abd-ElAzim |
| title_full_unstemmed | Cloning of genes from thermophilic bacteria and their expression in Bacillus subtilis and plants von Abd-ElAzim Farouk Abd-ElAzim |
| title_short | Cloning of genes from thermophilic bacteria and their expression in Bacillus subtilis and plants |
| title_sort | cloning of genes from thermophilic bacteria and their expression in bacillus subtilis and plants |
| topic | Proteasen (DE-588)4047521-9 gnd Analyse (DE-588)4122795-5 gnd Thermoactinomyces vulgaris (DE-588)4402189-6 gnd Heubacillus (DE-588)4242465-3 gnd Heterologe Genexpression (DE-588)4342015-1 gnd Xylose (DE-588)4190397-3 gnd Genklonierung (DE-588)4123275-6 gnd |
| topic_facet | Proteasen Analyse Thermoactinomyces vulgaris Heubacillus Heterologe Genexpression Xylose Genklonierung Hochschulschrift |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=007140455&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT abdelazimabdelazimfarouk cloningofgenesfromthermophilicbacteriaandtheirexpressioninbacillussubtilisandplants |