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.

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.

Further studies of L-arginine biosynthesis in germinating pea seeds. Evidence for the presence of argininosuccinate synthetase in cotyledon extracts

1969 ◽  
Vol 47 (4) ◽  
pp. 467-475 ◽  
Author(s):  
P. D. Shargool ◽  
E. A. Cossins
Keyword(s):  
Enzyme Activity ◽  
Gel Filtration ◽  
Magnesium Ions ◽  
Argininosuccinate Synthetase ◽  
Argininosuccinate Lyase ◽  
Specific Enzyme ◽  
Arginine Biosynthesis ◽  
Specific Enzyme Activity ◽  
Pea Seeds

The synthesis and metabolism of arginine in germinating peas was examined by supplying micromolar quantities of L-citruiline-carbamyl-14C, DL-arginine-carbamyl-14C, and DL-arginine-5-14C to imbibing seeds. Citrulline was readily incorporated into arginine, but the labelled arginine solutions were not extensively metabolized.Extracts of 1-day-old cotyledons were found to catalyze the synthesis of arginine from citrulline in a reaction having absolute requirements for ATP, L-aspartate, and magnesium ions. The extracts were fractionated by (NH4)2SO4 precipitation followed by gel filtration on columns of Sephadex G-50 and G-200. These treatments increased the specific enzyme activity by approximately 36 times. After such treatments the preparations still contained appreciable amounts of argininosuccinate lyase (L-argininosuccinate arginine-lyase, EC 4.3.2.1) activity. The rate of arginine synthesis was altered by increasing the concentrations of L-citrulline, L-aspartate, and ATP. The latter compounds were found to be inhibitory at concentrations of 1 μmole/ml and 4 μmoles/ml, respectively. Arginine synthesis was markedly affected by pH and by additions of arginine and argininosuccinate. It is concluded that germinating pea cotyledons contain appreciable levels of argirrinosuccmate synthetase (L-citrulline:L-aspartate ligase (AMP), EC 6.3.4.5), and furthermore, that this enzyme has importance in arginine biosynthesis during germination.

scholarly journals Fusaric Acid Production in Fusarium oxysporum Transformants Generated by Restriction Enzyme-Mediated Integration Procedure

2013 ◽  
Vol 19 (4) ◽  
pp. 254-258 ◽  
Author(s):  
Theresa Lee ◽  
Jean Young Shin ◽  
Seung Wan Son ◽  
Soohyung Lee ◽  
Jae-Gee Ryu
Keyword(s):  
Fusarium Oxysporum ◽  
Restriction Enzyme ◽  
Acid Production ◽  
Fusaric Acid ◽  
Integration Procedure

Isolation and some properties of argininosuccinate lyase from a higher plant source

1968 ◽  
Vol 46 (5) ◽  
pp. 393-399 ◽  
Author(s):  
P. D. Shargool ◽  
E. A. Cossins
Keyword(s):  
Enzyme Activity ◽  
Calcium Phosphate ◽  
Carboxymethyl Cellulose ◽  
Maximal Activity ◽  
Argininosuccinate Lyase ◽  
Arginine Biosynthesis ◽  
Isoelectric Precipitation ◽  
Higher Plant ◽  
Michaelis Constants ◽  
Pea Seeds

L-Argininosuccinate arginine-lyase (EC 4.3.2.1) was extracted from the cotyledons of germinating pea seeds. The enzyme was purified approximately 20 times by isoelectric precipitation, followed by treatment with calcium phosphate gel and chromatography on carboxymethyl cellulose. The partially purified enzyme was shown to catalyze both the condensation of arginine and fumarate to give argininosuccinate and the cleavage of argininosuccinate to give arginine and fumarate. The enzyme displayed maximal activity at pH 7.9 and was relatively stable after storage at −20 °C for 12 months or after dialysis overnight at 2 °C. The Michaelis constants for argininosuccinate and arginine were found to be 2 × 10−4 M and 6.7 × 10−3 M respectively. Enzyme activity was partially inhibited by 10−4 M p-chloromercuribenzoate. It is concluded that this enzyme has importance in arginine biosynthesis in germinating pea cotyledons.

Biochemical and genetic studies with arginine and proline auxotrophs of Neisseria gonorrhoeae

1989 ◽  
Vol 35 (12) ◽  
pp. 1069-1075 ◽  
Author(s):  
F. J. Picard ◽  
J. R. Dillon
Keyword(s):  
Neisseria Gonorrhoeae ◽  
The Other ◽  
Gene Defect ◽  
Proline Biosynthesis ◽  
Arginine Biosynthesis ◽  
Genetic Studies ◽  
Carbamoylphosphate Synthetase ◽  
Biosynthesis Gene ◽  
Gene Defects ◽  
Ornithine Transcarbamoylase

The prevalence of specific arginine biosynthesis gene defects was studied for 319 arginine-requiring clinical isolates of Neisseria gonorrhoeae by using the ability of the strains to utilize intermediates of arginine biosynthesis. Only 11 % of the uracil-requiring strains defective in the carbamylation of ornithine to yield citrulline had a defective carbamoylphosphate synthetase gene (carAB). Strains defective in carAB were of auxotype CUH. The other strains (89%) having a dual requirement for citrulline and uracil, which were mostly of auxotype PCU, were defective in the ornithine transcarbamoylase gene (argF). Over 90% of the strains were defective either in argJ (174 strains) or in argF (126 strains). Three argininosuccinate-requiring strains (i.e., defective in argG) of auxotype PAU were identified. Some of the arginine auxotrophs of N. gonorrhoeae defective in carAB, argJ, argF, or argG were complemented by genetic transformation with DNA from recombinant bacteriophages carrying characterized gonococcal arginine biosynthesis genes. Gene defects in proA (five strains) and in proB (six strains) were identified by gonococcal transformation assays with recombinant bacteriophages or plasmids carrying proline biosynthesis genes from N. gonorrhoeae. None of the 11 proline-requiring strains tested was defective in proC.Key words: Neisseria gonorrhoeae, arginine biosynthesis, proline biosynthesis, gene defect, marker rescue.

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