Promoter-tagged restriction enzyme-mediated insertion (PT-REMI) mutagenesis in

1999 ◽  
Vol 262 (1) ◽  
pp. 27 ◽  
Author(s):  
J. R. Shuster ◽  
M. Bindel Connelley
Keyword(s):  
Restriction Enzyme ◽  
Remi Mutagenesis

Mutagenesis via Insertional or Restriction Enzyme-Mediated Integration (REMI) as a Tool to Tag Pathogenicity Related Genes in Plant Pathogenic Fungi

1999 ◽  
Vol 380 (7-8) ◽  
pp. 855-864 ◽  
Author(s):  
F. J. Maier ◽  
W. Schäfer
Keyword(s):  
Restriction Enzyme ◽  
Molecular Basis ◽  
Insertional Mutagenesis ◽  
Transformation Efficiency ◽  
Pathogenic Fungi ◽  
Transformation Protocol ◽  
Vector Integration ◽  
Future Developments ◽  
Genetically Determined ◽  
Remi Mutagenesis

Abstract Random insertional mutagenesis is a powerful tool to investigate the molecular basis of most genetically determined processes, for example in pathogenic fungi. An improved version of this method is the insertional mutagenesis via restriction enzyme mediated integration (REMI). Transformation efficiency and mode of vector integration are species dependent and further influenced by vector conformation, restriction enzyme activity, and transformation protocol. An overview is given, covering the mutants and already identified genes obtained after REMI mutagenesis. An outlook describes the future developments in the field.

scholarly journals Mutation of an Arginine Biosynthesis Gene Causes Reduced Pathogenicity in Fusarium oxysporum f. sp. melonis

2001 ◽  
Vol 14 (4) ◽  
pp. 580-584 ◽  
Author(s):  
Fumio Namiki ◽  
Michiko Matsunaga ◽  
Mitsuru Okuda ◽  
Iori Inoue ◽  
Kazufumi Nishi ◽  
...  
Keyword(s):  
Structural Analysis ◽  
Fusarium Oxysporum ◽  
Restriction Enzyme ◽  
Argininosuccinate Lyase ◽  
Arginine Biosynthesis ◽  
Biosynthesis Gene ◽  
Remi Mutagenesis ◽  
Auxotrophic Mutation ◽  
Arginine Auxotroph

Restriction enzyme-mediated integration (REMI) mutagenesis was used to tag genes required for pathogenicity of Fusarium oxysporum f. sp. melonis. Of the 1,129 REMI transformants tested, 13 showed reduced pathogenicity on susceptible melon cultivars. One of the mutants, FMMP95–1, was an arginine auxotroph. Structural analysis of the tagged site in FMMP95-1 identified a gene, designated ARG1, which possibly encodes argininosuccinate lyase, catalyzing the last step for arginine biosynthesis. Complementation of FMMP95–1 with the ARG1 gene caused a recovery in pathogenicity, indicating that arginine auxotrophic mutation causes reduced pathogenicity in this pathogen.

Sign in / Sign up

Export Citation Format

Share Document