Methods for investigating nucleo-cytoplasmic transport of RNA: a laboratory manual
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| Main Authors: | , , |
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| Format: | Book |
| Language: | English |
| Published: |
Stuttgart u.a.
Fischer
1989
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| Subjects: | |
| Online Access: | Inhaltsverzeichnis |
| Item Description: | Literaturverz. S. [147] - 156 |
| Physical Description: | 156 S. Ill., graph. Darst. |
| ISBN: | 3437112619 0895742969 |
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Record in the Search Index
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| adam_text | IMAGE 1
METHODS FOR INVESTIGATING
NUCLEO-CYTOPLASMIC TRANSPORT OF RNA A LABORATORY MANUAL
HEINZ C._SCHR6DER, MICHAEL BACHMANN AND WERNER E. G. MIILLER
WITH 42 FIGURES, 3 IN COLOUR
GUSTAV FISCHER VERLAG * STUTTGART * NEW YORK * 1989
IMAGE 2
CONTENTS
ABBREVIATIONS 7
I. INTRODUCTION 9
II. PREPARATION OF NUCLEI AND NUCLEAR SUBSTRUCTURES 15
A) NUCLEI 15
B) NUCLEAR MATRICES 18
C) NUCLEAR ENVELOPES 20
D) DEMEMBRANATED NUCLEI *. 24
E) NUCLEAR GHOSTS 25
F) PORE-COMPLEX LAMINAE 25
G) NUCLEAR ENVELOPE VESICLES (RESEALED VESICLES) 26
III. PURIFICATION OF COMPONENTS OF THE MRNA TRANSLOCATION SYSTEM 28 A)
NUCLEAR ENVELOPE NTPASE 28
B) POLY(A) RECOGNIZING CARRIER PROTEIN 32
C) MRNA TRANSPORT STIMULATORY PROTEINS 34
D) NUCLEAR ENVELOPE-ASSOCIATED NL-LIKE AND NIL-LIKE PROTEIN KINASES 37
IV. PURIFICATION OF PROTEINS AND ENZYMES RELATED TO RNA TRANSPORT 40
A) POLY(A) POLYMERASE-ENDORIBONUCLEASE IV COMPLEX 40
B) ENDORIBONUCLEASE IV AND POLY(A) POLYMERASE 42
C) 2 , 3 -EXORIBONUCLEASE 45
D) PROTEIN KINASE C 49
E) LA AND RO PROTEIN 52
V. ISOLATION OF RNA AND RNP 55
A) TOTAL RNA 55
B) POLY(A) (+) RNA 57
C) POLYSOMES 58
D) POLYSOMAL POLY(A) (+) MRNA 58
E) PREPARATION OF POLY(A) (-) MRNA 60
F) PREPARATION OF THE PO!Y(A)-RNP COMPLEX 60
G) PREPARATION OF THE POLY(A) COMPONENT OF THE POLY(A)-RNP COMPLEX 61 H)
SNRNA T 62
I) TRNA 63
VI. RNA TRANSPORT MEASUREMENTS 66
A) IN VITRO SYSTEM FOR MEASUREMENT OF RNA RELEASE 66
B) IN VITRO SYSTEM FOR MEASUREMENT OF RNA TRANSLOCATION 69
C) IN VITRO NUCLEAR TRANSCRIPTION/TRANSPORT MEASUREMENTS 73
D) RNA INFLUX AND EFFLUX MEASUREMENTS USING NUCLEAR ENVELOPE VESICLES 75
IMAGE 3
6 CONTENTS
VII. ENZYME ASSAYS 77
A) NTPASE 77
B) PROTEIN KINASES NI AND NIL 80
C) PROTEIN KINASE C 81
D) PHOSPHOPROTEIN PHOSPHATASE 82
E) POLY(A) POLYMERASE 83
F) ENDORIBONUCLEASE IV 86
G) 2 , 3 -EXORIBONUCLEASE 87
H) TOPOISOMERASES I AND II 87
VIII. DETECTION OF RNA 90
A) NORTHERN-BLOT HYBRIDIZATION 90
B) DOT-BLOT HYBRIDIZATION 93
C) HYBRIDIZATION TO POLY(A) (+) RNA BOUND TO POLY(U) PAPER 94
D) SPOT HYBRIDIZATION 94
E) IN SITU HYBRIDIZATION 96
IX. PREPARATION OF DNA PROBES 99
A) TRANSFORMATION OF BACTERIA BY PLASMID DNA (CACK PROCEDURE) 99 B)
MASTER PLATE 100
C) PERMANENT GLYCEROL CULTURE 100
D) PLASMID PREPARATION 100
E) PURIFICATION OF PLASMID DNA 102
F) RESTRICTION ANALYSIS 103
G) AGAROSE GEL ELECTROPHORESIS 104
H) PREPARATION OF BACTERIA FOR PHAGE CULTURE 105
I) PLATING OF LAMBDA PHAGES 106
J) PREPARATION OF BACTERIOPHAGE LAMBDA DNA 107
K) NICK TRANSLATION 110
1) LABELLING AT 3 END OF RNA I LL
X. ANALYSIS OF RNA 112
A) GEL ELECTROPHORESIS OF RNA 112
B) SUCROSE GRADIENT CENTRIFUGATION 113
C) SEQUENCING OF RNA 114
XI. ANALYSIS OF PROTEIN 118
A) SDS POLYACRYLAMIDE GEL ELECTROPHORESIS 118
B) DETERMINATION OF INCORPORATION OF PHOSPHATE 119
XII. BINDING STUDIES 121
A) BINDING OF POLY(A) TO PORE-COMPLEX LAMINAE 121
B) BINDING OF IJDBU 123
XIII. FRACTIONATION OF NUCLEAR SUBSTRUCTURES 125
A) SOLUBILIZATION OF THE NUCLEAR ENVELOPE POLY(A ) BINDING SITE 125
B) EXTRACTION OF NUCLEAR ENVELOPES WITH TRITON X-114 125
XIV. PHOTOAFFINITY LABELLING OF NUCLEAR MATRIX AND NUCLEAR ENVELOPE
PROTEINS BY 8-AZIDO-ATP 129
IMAGE 4
ABBREVIATIONS * 7
XV. IMMUNOLOGICAL METHODS 130
A) PRODUCTION OF MONOCLONAL ANTIBODIES 130
B) ENZYME-LINKED IMMUNOSORBENT ASSAY (ELISA) 130
C) WESTERN BLOTTING 131
D) IMMUNOPRECIPITATION 132
XVI. IMMUNOFLUORESCENCE MICROSCOPY 133
XVII. MICROINJECTION 140
XVIII. IN VITRO TRANSLATION OF TRANSPORTED RNA 143
XIX. ELECTRON MICROSCOPY 144
ACKNOWLEDGEMENTS 146
REFERENCES 147
ABBREVIATIONS
A, P-METHYLENE ATP, ADENOSINE 5 -( X, P-METHYLENE)TRIPHOSPHATE P,
Y-METHYLENE ATP, ADENOSINE 5 -(P, Y-METHYLENE)TRIPHOSPHATE CDNA,
COMPLEMENTARY DNA CPM, COUNTS PER MINUTE
DEAE, DIETHYLAMINOETHYL DMAPN, 3-DIMETHYLAMINOPROPIONITRILE DMSO,
DIMETHYL SULPHOXIDE DPM, DISINTEGRATIONS PER MINUTE EDTA,
ETHYLENEDIAMINETETRAACETIC ACID EGTA, ETHYLENEGLYCOL BIS(P-AMINOETHYL
ETHER)-N, N, N , N -TETRAACETIC ACID ELISA, ENZYME-LINKED IMMUNOSORBENT
ASSAY FITC, FLUORESCEIN ISOTHIOCYANATE
HEPES, N-2-HYDROXYETHYLPIPERAZINE-N -2-ETHANESULFONIC ACID HIV-1, HUMAN
IMMUNODEFICIENCY VIRUS TYPE 1 HNRNA, HETEROGENEOUS NUCLEAR RNA HSV-1,
HERPES SIMPLEX VIRUS TYPE 1 KB, KILOBASES KDA, KILODALTON MAB,
MONOCLONAL ANTIBODY M-AMSA, 4 -(9-ACRIDINYLAMINO 1
)METHANESULFBN-M-ANISIDIDE MOPS, 3-[N-MORPHOLINO]PROPANESULFONIC ACID
MRNA, MESSENGER RNA NTPASE, NUCLEOSIDE TRIPHOSPHATASE O.D., OPTICAL
DENSITY PDBU, PHORBOL-12,13-DIBUTYRATE
P.I., POST INFECTION
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| indexdate | 2024-07-09T17:07:24Z |
| institution | BVB |
| isbn | 3437112619 0895742969 |
| language | English |
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| physical | 156 S. Ill., graph. Darst. |
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| spelling | Schröder, Heinz C. 1951- Verfasser (DE-588)120765039 aut Methods for investigating nucleo-cytoplasmic transport of RNA a laboratory manual Heinz C. Schröder, Michael Bachmann and Werner E. G. Müller Stuttgart u.a. Fischer 1989 156 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Literaturverz. S. [147] - 156 Biological Transport Laboratory Manuals Cytoplasm metabolism Laboratory Manuals Messenger RNA Physiological transport Laboratory manuals RNA metabolism Laboratory Manuals RNS (DE-588)4076759-0 gnd rswk-swf Transport (DE-588)4060680-6 gnd rswk-swf RNS (DE-588)4076759-0 s Transport (DE-588)4060680-6 s DE-604 Bachmann, Michael Verfasser aut Müller, Werner E. G. 1942- Verfasser (DE-588)115475893 aut HEBIS Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=005034274&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Schröder, Heinz C. 1951- Bachmann, Michael Müller, Werner E. G. 1942- Methods for investigating nucleo-cytoplasmic transport of RNA a laboratory manual Biological Transport Laboratory Manuals Cytoplasm metabolism Laboratory Manuals Messenger RNA Physiological transport Laboratory manuals RNA metabolism Laboratory Manuals RNS (DE-588)4076759-0 gnd Transport (DE-588)4060680-6 gnd |
| subject_GND | (DE-588)4076759-0 (DE-588)4060680-6 |
| title | Methods for investigating nucleo-cytoplasmic transport of RNA a laboratory manual |
| title_auth | Methods for investigating nucleo-cytoplasmic transport of RNA a laboratory manual |
| title_exact_search | Methods for investigating nucleo-cytoplasmic transport of RNA a laboratory manual |
| title_full | Methods for investigating nucleo-cytoplasmic transport of RNA a laboratory manual Heinz C. Schröder, Michael Bachmann and Werner E. G. Müller |
| title_fullStr | Methods for investigating nucleo-cytoplasmic transport of RNA a laboratory manual Heinz C. Schröder, Michael Bachmann and Werner E. G. Müller |
| title_full_unstemmed | Methods for investigating nucleo-cytoplasmic transport of RNA a laboratory manual Heinz C. Schröder, Michael Bachmann and Werner E. G. Müller |
| title_short | Methods for investigating nucleo-cytoplasmic transport of RNA |
| title_sort | methods for investigating nucleo cytoplasmic transport of rna a laboratory manual |
| title_sub | a laboratory manual |
| topic | Biological Transport Laboratory Manuals Cytoplasm metabolism Laboratory Manuals Messenger RNA Physiological transport Laboratory manuals RNA metabolism Laboratory Manuals RNS (DE-588)4076759-0 gnd Transport (DE-588)4060680-6 gnd |
| topic_facet | Biological Transport Laboratory Manuals Cytoplasm metabolism Laboratory Manuals Messenger RNA Physiological transport Laboratory manuals RNA metabolism Laboratory Manuals RNS Transport |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=005034274&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
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