Journal: Scientific Reports

Article Title: Down-regulation of TUFM impairs host cell interaction and virulence by Paracoccidioides brasiliensis

doi: 10.1038/s41598-019-51540-y

Figure Lengend Snippet: Obtaining P. brasiliensis PbTufM aRNA isolate. ( A ) Agarose gel electrophoresis to confirm the transference of HPH gene by ATMT to PbEV (PbWT transformed with empty vector) and PbTufM aRNA (PCR amplification yielded a 1,000 bp internal fragment), NC: negative control of PCR reaction, MW: 1Kb DNA ladder (GeneDireX). ( B ) Expression levels of PbTUFM assessed by Real-Time PCR in PbWT, PbEV, and PbTufM aRNA yeast cells after being subcultured for 1 and 12 months (gene expression levels obtained by RT-PCR were normalized to the level of expression of the internal reference, β-tubulin; *** p < 0.0001 compared with PbWT and PbEV.

Article Snippet: To obtain an isolate with reduced PbTUFM gene expression, we used aRNA and ATMT according to the methods described by Almeida et al . . DNA from Pb18 was isolated and the fragment referring to aRNA oligonucleotide targeting the exon coding sequence of PbTUFM (PADG_01949.1; Broad Institute) amplified using a high-fidelity proofreading DNA polymerase .

Techniques: Agarose Gel Electrophoresis, Transformation Assay, Plasmid Preparation, Amplification, Negative Control, Expressing, Real-time Polymerase Chain Reaction, Gene Expression, Reverse Transcription Polymerase Chain Reaction

Journal: Scientific Reports

Article Title: Down-regulation of TUFM impairs host cell interaction and virulence by Paracoccidioides brasiliensis

doi: 10.1038/s41598-019-51540-y

Figure Lengend Snippet: Primers designed for antisense oligonucleotide and for hph PCR amplification, and for RT-qPCR analyses.

Article Snippet: To obtain an isolate with reduced PbTUFM gene expression, we used aRNA and ATMT according to the methods described by Almeida et al . . DNA from Pb18 was isolated and the fragment referring to aRNA oligonucleotide targeting the exon coding sequence of PbTUFM (PADG_01949.1; Broad Institute) amplified using a high-fidelity proofreading DNA polymerase .

Techniques: Amplification