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Several seroconversions occurring in 2002 among sentinel cattle during the bluetongue-vaccination campaign in Lazio and Tuscany (central Italy) led to the suspicion of vaccine-virus circulation. Therefore in 2003, 17 seroconverting sentinel herds were investigated for the characteristics of the virus involved. From these farms, 91 unvaccinated animals and 57 Culicoides pools were tested for the presence of the bluetongue vaccine virus (serotype-2) or other strains. The presence of vaccine virus serotype-2 was confirmed by PCR followed by restriction analysis in the whole blood of 17 unvaccinated sentinel cattle and 12 pools of Culicoides imicola or C. obsoletus. Of the 17 herds, five were positive only for vaccine virus serotype-2, four were positive for other strains and two for both the vaccine and other strains; the remaining premises were virologicaly negative. The vaccine virus serotype-2 also was detected in areas not included in the vaccination campaign.

The nucleotide (nt) sequence of the dihydrofolate reductase (DHFR)-encoding gene (DFRl) of Candida albicans was determined. The gene contains an open reading frame of 576 nt, coding for a protein of 192 amino acid (aa) residues (calculated Mr. 22222), that is 38.5 and 31% similar to the Saccharomyces cerevisiae and human enzymes, respectively. The first 36 residues, at the N terminus, of the deduced aa sequence are identical to those determined by sequencing of the purified enzyme from C. albicans. Putative transcription start points were also determined. Restriction-fragmentlength polymorphism analysis of the DFR1 chromosomal region suggests the presence of a single copy of the gene per haploid genome and shows a limited variability among the different C. albicans strains tested.

In bacteria the initiation of protein synthesis is a complex phenomenon in which specific proteins, termed initiation factors (IFs) IF1, IF2, and IF3, are involved. Notwithstanding the progress made in understanding their functions, the precise molecular mechanisms of action of these factors remain somewhat obscure. One reason for this lack of knowledge is the difficulty involved in purifying sufficient quantities of these proteins. We have developed a new procedure for purification of IFs from recombinant Escherichia coli strains producing high levels of E. coli IF3 and Bacillus stearothermophilus IF2. This new procedure is quicker than previous methods, easily scaled up to large volumes, and can be used, with only minor modifications, for different IFs. This new purification method consists essentially of one chromatographic (FPLC) separation on an ion-exchange resin (S-Sepharose fast-flow or Mono-S HR). Using this procedure we have been able to obtain chromatographically pure and biologically active preparations of both IF2 and IF3.