The gene (pox1) encoding a phenol oxidase from Pleurotus ostreatus, a lignin-degrading basidiomycete, was cloned and sequenced, and the corresponding pox1 cDNA was also synthesized and sequenced. The isolated gene consists of 2,592 bp, with the coding sequence being interrupted by 19 introns and flanked by an upstream region in which putative CAAT and TATA consensus sequences could be identified at positions ؊174 and ؊84, respectively. The isolation of a second cDNA (pox2 cDNA), showing 84% similarity, and of the corresponding truncated genomic clones demonstrated the existence of a multigene family coding for isoforms of laccase in P. ostreatus. PCR amplifications of specific regions on the DNA of isolated monokaryons proved that the two genes are not allelic forms. The POX1 amino acid sequence deduced was compared with those of other known laccases from different fungi.
High-efficiency transformation of the autogamous diploid legume Lotus japonicus by means of Agrobacterium rhizogenes was used to develop plant lines expressing a promoter-less gusA gene in a nodulation- or lateral root-associated manner. The approach exploits the putatively preferential integration of T-DNA into actively transcribed regions, thereby providing an enrichment for gene tagging events associated with the quickly assayable activation of a gusA promoter-less construct. Taking advantage of this enrichment and selection strategy, a T-DNA tagging program was initiated and screening for β-glucuronidase (GUS) activity was performed on root clones isolated after transformation with a gusA-promoter-less binary vector. The aim of this approach is the identification of genes involved in nodule formation induced by Mesorhizobium loti, lateral root organogenesis, and the eventual isolation of corresponding mutants. A large collection (220) of GUS-positive transformants showing a variety of expression patterns in different regions of roots and nodules was obtained; a preliminary molecular characterization of these plants is presented.
Background: Heat and cold shock response are normally considered as independent phenomena. A small amount of evidence suggests instead that interactions may exist between them in two Lactococcus strains.
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