Transposon tagging is a very useful tool for insertional mutagenesis and gene cloning. Transposons can be employed to tag genes both in Procaryota and Eucaryota. The aim of this project is to obtain an A. nidulans mutant with a suppression of proline auxotrophy by using a heterologous transposon Impala from Fusarium oxysporum. A.nidulans initially strain was a mutant in proA gene, thus it had a pro- phenotype. Using transposon mutagenesis we obtained a suX(pro) mutant which, in spite of mutation in proA gene, was able to grow on a medium without proline. Inverse PCR method was used for amplification of tagged gene. The suX(pro) gene was amplified together with upstream and downstream regions. Further it was cloned on plasmid vector and sequenced. An open reading frame of 1335bp was identified. It contains one putative intron (92bp) into which the Impala element has been transposed. The putative product of the suX(pro) gene is a protein of 259 amino acids residues. This protein comprises two conservative RRM domains (RNA recognition motif), each of them contains two RNP-SC regions (ribonucleoprotein consensus sequence). Proteins comprising RRM domains play a crucial role in posttranscriptional regulation of gene expression at the level of poliadenylation, splicing, transport of mRNAs from nucleus to cytoplasm and in mRNAs stabilization.
Fungal Genet. Newsl. 50 (Supl):abstract
Full conference title:
22nd Fungal Genetics Conference
- Fungal Genetics Conference 22nd (2001)