Abstract

The genetic polymorphism of Paracoccidioides spp. isolates has been widely demonstrated by different authors using several molecular techniques. The effects of transposable elements (TEs) may be responsible for the genetic polimorphism observed in fungi genome. Recently, eight new Tc1/mariner families referred as Trem (Transposable element mariner) labeled A through H were identified in Paracoccidioides spp. genome. The distribution of Trem elements varied between the genomic sequences characterized as belonging to P. brasiliensis (S1 and PS2) and P. lutzii(new species proposed to comprise Pb01-like isolates): TremC and TremH were found in all species; TremB,F and G in the phylogenetic species S1 and PS2; TremD only in S1 and TremE exclusively in P. lutzii. This study aims to investigate the ocurrence of Trem A-H in the phylogenetic species 3 (PS3) isolates not studied before.DNA of two PS3 isolates (EPM83 and 77) was extracted and subsequently subjected to Polymerase Chain Reaction (PCR) of terminal inverted repeats (TIRs) of each Trem, using primers previously described. PCR were carried out on 10 ng of DNA in a 20 μL reaction mixture containing 10 mM Tris-Hcl pH 9, 1.5 mM MgCl2, 50 mM KCl, 100 mM of each dNTP, 100 pmol of each oligonucleotide and 1 unit of Taq DNA Polimerase. The cycling conditions were as follows: denaturing at 94&bullC for 5 min, annealing at the Tm for each primer for 30 s and extension at 72&bullC for 1 min. At the end of the 25th cycle, the heat-denaturing step was omitted and extension was allowed to proceed at 72&bullC for 5 min. The amplified product was visualized on 1% agarose gel stained with ethidium bromide.All the elements, with the exception of TremH were amplified in both isolates.The strategy developed in the present study proved effective in identifying Trem elements in both PS3 isolates. The non-amplification of TremE supports the hypothesis that this element is present only in Pb01-like isolates. It would be interesting to verify the ocurrence of Trem elements in a larger number of isolates in such a way to better understand the pattern distribution of these elements in the phylogenetic species 3.਀㰀⼀昀漀渀琀㸀㰀⼀瀀㸀㰀戀爀㸀㰀戀㸀䬀攀礀眀漀爀搀㨀 㰀⼀戀㸀☀渀戀猀瀀㬀䐀一䄀 琀爀愀渀猀瀀漀猀漀渀猀Ⰰ 倀栀礀氀漀最攀渀攀琀椀挀 猀瀀攀挀椀攀猀 ㌀Ⰰ 吀挀㄀⼀洀愀爀椀渀攀爀Ⰰ 吀爀攀洀 䄀ⴀ䠀㰀⼀琀搀㸀㰀⼀琀爀㸀㰀⼀琀愀戀氀攀㸀㰀⼀琀爀㸀㰀⼀琀搀㸀㰀⼀琀愀戀氀攀㸀㰀⼀戀漀搀礀㸀㰀⼀栀琀洀氀㸀