Objective
The objective of this research is to measure experimentally the frequency of horizontal gene transfer between eukaryotic organisms. The model system we have chosen involves the plant pathogen Cladosporium fulvum and its host species, tomato. The genes that will be studied will be plant and fungal retrotransposons, as we consider these genes are best equipped to invade non-host genomes.
An assessment of the risks involved in the release of genetically engineered organisms must take into account the frequency of horizontal gene transfer. The frequency of horizontal gene transfer places an upper limit on the safety of the deliberate release of engineered organisms because it will indicate the likelihood of gene escape to unrelated species in the environment. The observation that an element related to the C fulvum retrotransposon CfT1 is present in tomato may indicate recent interspecies genetic exchange. The objective of this research is to experimentally measure the frequency of horizontal gene transfer between eukaryotic organisms.
Initial objectives are;
the creation and testing in C fulvum and tomato of chimeric marker genes;
the creation of marked copies of both transposons and transformation into C fulvum and tomato;
collection of worldwide Solanaceae strains;
screening of the plants by hybridisation polymerase chain reaction (PCR) and sequencing;
collection of worldwide C fulvum strains;
screening of the C fulvum strains by hybridisation PCR, and sequencing.
Results to date include;
chimeric marker genes have been created in both laboratories;
the creation of marked copies of both transposons is under way;
a Tnt1 LTR-GUS construct has been shown to be specifically activated by cell wall hydrolases;
31 C fulvum strains have been collected worldwide and have been screened by hybridisation.
Tobacco, tomato and C. fulvum all contain retrotransposons in their genomes. These elements have no known role beneficial to their hosts; their sequence homology and distribution provides strong circumstantial evidence of horizontal gene transfer. The rationale behind this project is to establish the situation in which horizontal gene transfer might take place and to be able to detect such an event even if it is very rare. The first condition is provided by the intimate relationship between a biotrophic plant pathogen and its host plant. The second condition is provided by the creation of test genes whose transfer can be detected even amongst a vast excess of cells lacking the transferred genes.
Engineered copies of the retrotransposon Cft-1, from C. fulvum, will be created by including chimeric genes for either hydromycin phosphotransferase or beta-glucuronidase using either the CaMV 35S or A. nidulans gpd promoters. Similarly, copies of the tobacco retrotransposon Tnt-1 will be created with the chimeric genes. Control experiments will establish whether the chimeric genes are active in both genomes. Next, the marked transposons will be transformed into both C. fulvum, and tomato. Ultimately marked C. fulvum strains will be infected into tomato and plant cells carrying the marker gene will be selected. Conversely, marked strains of tomato will be infected With wild type C. fulvum. Fungal spores, from the infected leaves, carrying the marker gene will be sought.
Fields of science (EuroSciVoc)
CORDIS classifies projects with EuroSciVoc, a multilingual taxonomy of fields of science, through a semi-automatic process based on NLP techniques. See: The European Science Vocabulary.
CORDIS classifies projects with EuroSciVoc, a multilingual taxonomy of fields of science, through a semi-automatic process based on NLP techniques. See: The European Science Vocabulary.
- medical and health sciences medical biotechnology genetic engineering gene therapy
- natural sciences biological sciences genetics genomes
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Coordinator
NR4 7TJ Norwich
United Kingdom
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