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Language(s):
eng - English
Dates:
2017-08-092017-08-09
Publication Status:
Issued
Pages:
-
Publishing info:
Kiel : ANY PUBLISHER
Table of Contents:
Contents
Introduction ................................ ................................ ................................ .......................... 2
Plant pathogen interactions ......................................................................................................... 2
Fungal pathogens ........................................................................................................................ 3
Zymoseptoria tritici ...................................................................................................................... 4
Aim .............................................................................................................................................. 6
Methods and Material ................................ ................................ ................................ .......... 7
Creation of a deletion mutant ....................................................................................................... 7
Enzymes and other materials ................................................................................................... 7
Oligonucleotides and plasmids................................................................................................. 7
Polymerase chain reaction ....................................................................................................... 9
Gel electrophoresis and isolation of the fragments ................................................................... 9
Gibson assembly ..................................................................................................................... 9
Overlap PCR .......................................................................................................................... 10
Transformation of E. coli ........................................................................................................ 11
Plasmid-DNA Isolation ........................................................................................................... 11
Verification of the plasmid ...................................................................................................... 12
Agrobacterium mediated transformation of Z. tritici ................................................................ 12
DNA isolation and verification of the mutants with PCR ......................................................... 13
DNA Isolation for the Southern blot ........................................................................................ 14
Southern blot for further verification ....................................................................................... 14
Plant experiment ....................................................................................................................... 15
Results ................................ ................................ ................................ ............................... 17
Creation of the deletion mutant .................................................................................................. 17
Amplification of the plasmid’s fragments ................................................................................ 17
Assembly of the vector and transformation of E. coli .............................................................. 18
Transformation of Z. tritici ...................................................................................................... 20
Plant experiment ....................................................................................................................... 23
Manual evaluation .................................................................................................................. 23
Analysis with ImageJ ............................................................................................................. 25
Combined data ...................................................................................................................... 28
Discussion ................................ ................................ ................................ ......................... 29
Zusammenfassung ................................ ................................ ................................ ............ 32
References ................................ ................................ ................................ ........................ 33
Acknowledgements ................................ ................................ ................................ ............ 35
Supplement ................................ ................................ ................................ ........................ 36
Erklärung ................................ ................................ ................................ ........................... 41
Rev. Type:
-
Identifiers:
Other: Dipl/12915
Degree:
Bachelor
