Development of a cauliflower in vitro editing system and analysis of various cis- and trans-factors [Elektronische Ressource] / vorgelegt von Johannes Alexander van der Merwe

Dre Van Der Merwe

Sujets

• Biologie

Informations

• Publié par: universitat_ulm
• Publié le: 01 janvier 2007
• Nombre de lectures: 22
• Langue: English
• Poids de l'ouvrage: 2 Mo

Extrait

Abteilung Molekulare Botanik

(Leiter Prof.Dr. Axel Brennicke)

Universität Ulm

Development of a cauliflowerin vitroediting system and analysis of variouscis-and-trans-factors

Dissertation

zur Erlangung des Doktorgrades (Dr. rer. nat.)

an der Fakultät für Naturwissenschaften

der Universität Ulm

vorgelegt von

Johannes Andreas van der Merwe

aus Johannesburg

2007

Amtierender Dekan der Fakultät für Naturwissenschaften:

Prof. Dr. Klaus-Dieter Spindler

Erstgutachter:

Prof. Dr. Axel Brennicke, Abteilung Molekulare Botanik, Universität Ulm

Zweitgutachter: Dr. Anita Marchfelder

Datum der Promotion:

Die Arbeiten im Rahmen der vorgelegten Dissertation wurden in der Abteilung Molekulare Botanik der Universität Ulm durchgeführt und von Herrn Prof. Dr. Axel Brennicke betreut.

Ulm, den 10.5.2007

3.12

Isolating specific RNA binding proteins using glass beads.......................................

18

Zusammenfassung.............................................................................................................2

1   Summary..........................................................................................................................1

Table of Contents

i

15

3.9

15

 In-vitro RNA editing reactions...................................................................................

3.10

Detection of the RNA editing activity by mismatch analysis......................................

16

3.11

 RNA substrates........................................................................................................

3.8

15

 Preparation of mitochondrial extracts.......................................................................

3.7

The extraction of plant mitochondria.........................................................................

14

3.6

13

 Standard methods in molecular biology....................................................................

3.5

13

 Devices....................................................................................................................

3.4

13

 Oligonucleotides ......................................................................................................

12

3.3

2   Introduction.....................................................................................................................3

 Enzymes..................................................................................................................

2.1

 RNA editing: an overview...........................................................................................3

2.2

 Methods and effects of RNA editing...........................................................................3

2.3

 RNA editing in trypanosomes ....................................................................................6

2.4

 RNA editing in animals...............................................................................................7

RNA editing in plants: plastids and mitochondria ....................................................... 8

2.5

2.6

Aims of this study .................................................................................................... 11

 Plant material...........................................................................................................12

3.2

3 Materials and Methods.................................................................................................12

3.1

 Consumables...........................................................................................................

12

4   Results...........................................................................................................................20

4.1

4.2

4.3

4.4

 Screening for cis-elements in silico..........................................................................20

 Optimising the pea in vitro system............................................................................20

Characterising cis-elements in the 2nd editing site of atp9....................................... 22

 Investigating possible trans-acting elements............................................................26

Is RNA involved in RNA editing in higher plants?.............................................................26

The involvement of a transaminase?................................................................................29

Energy requirements of the in vitro reaction..................................................................... 31

Differences in editing sites between different Arabidopsis ecotypes ................................ 33

5   Discussion and Conclusions ......................................................................................35

Determination of cis elements in silico..............................................................................35

Development of the cauliflower in vitro editing system.....................................................35

Cis-elements for the 2nd editing site in the atp9 mRNA transcript.................................... 36

Is external RNA involved in RNA editing in plant mitochondria?....................................... 37

The involvement of a deaminase or transaminase?.........................................................38

Energy requirements of the in vitro reaction..................................................................... 39

Differences in editing sites between different Arabidopsis ecotypes................................. 40

6   References.....................................................................................................................41

7   Appendix........................................................................................................................49

7.1

 Publications..............................................................................................................

49

ii