An autonomously replicating plasmid transforms Aspergillus nidulans at high frequency

Comparison between polyethylene glycol- and polyethylenimine-mediated transformation of Aspergillus nidulans

“…By including an autonomously replicating sequence (ARS), AMA1, in the transforming plasmid pTN51, it was possible to avoid chromosomal integration. Presence of this sequence confers a 250-fold difference in transformation frequency (Gems et al 1991). The major steps involved in transformation with the ARS plasmid include passage through the plasma membrane, cytoplasm, and nuclear envelope, expression of the marker gene, and replication of the DNA.…”

Inducer-Dependent Nuclear Localization of a Zn(II)2Cys6Transcriptional Activator, AmyR, inAspergillus nidulans

“…Then the BsrG I and EcoR I digested amyR fragment (2.5-kb) was introduced between the BsrG I and EcoR I sites of the resulting plasmid so that the amyR gene was fused in-frame to the C-terminus of the GFP gene. A 2.6-kb Pst IBamH I fragment carrying the A. nidulans argB gene, derived from pDHG25, 34) was inserted at the corresponding sites of pBluescript II KS(+) to construct pAR5. The alcAp-GFP-AmyR fusion gene described above was recovered as the Kpn I-EcoR I fragment (3.8-kb) and was introduced between the Kpn I and EcoR I sites of pAR5, resulting in the plasmid pAGAR-F.…”

Evidence for Dicer-dependent RNA interference in the industrial penicillin producer Penicillium chrysogenum

“…pPTRII (Supplementary Table S2), carrying the AMA1-based elements together with the pyrithiamin-resistance gene ptrA from Aspergillus oryzae (Gems et al, 1991;Kubodera et al, 2000Kubodera et al, , 2002, was modified by introducing complementary annealed oligonucleotides (Nhe_1 and Nhe_2) into the KpnI-HindIII restriction sites. This construction generated a single NheI site in plasmid pPTRII-NheI.…”