Cloning, in silico structural characterization and expression analysis of MfAtr4, an ABC transporter from the banana pathogen Mycosphaerella fijiensis

ABC transporters are membrane proteins that use the energy released from the hydrolysis of ATP to drive the transport of compounds across biological membranes. In some plants, pathogenic fungi ABC transporters play a role as virulence factors by mediating the export of plant defense compounds or fungal virulence factors. Mycosphaerella fijiensis, the causal agent of black Sigatoka disease in banana, is the main constraint for the banana industry worldwide. So far, little is known about molecular mechanism that it uses to infect the host. In this study, degenerated primers designed from fungal ABC transporters known to be involved in virulence were used to isolate homologs from M. fijiensis. Here, we reported the full cloning of MfAtr4 a putative ortholog of MgAtr4, an ABC transporter of the related Mycosphaerella graminicola with a function in virulence. Similarities and differences with its presumed ortholog MgAtr4 are described, and the putative function of MfAtr4 are discussed. Analysis of MfAtr4 gene expression in field banana samples exhibiting visible symptoms of black Sigatoka disease indicated a higher expression of MfAtr4 during the first symptomatic stages in comparison to the late necrotrophic phases, suggesting a role for MfAtr4 in the early stages of pathogenic development of M. fijiensis.Key words: ABC transporters, virulence factors, MgAtr4 ortholog, Mycosphaerella fijiensis, black Sigatoka, Musa sp

Design of qPCR primers for the early region of Human Papillomavirus oncogenic types 16 and 18

Primer pairs for the HR-HPV16 and 18 were designed to target the open reading frames (ORF) E6, E7, E1 and E2 in real-time PCR assays. This data is provided in the folders: Figures, Tables and HPV Alignments 1. HPV alignments folder contain the following files: For gene-specific qPCR primer design, poorly conserved nucleotide regions were identified from HPV alignments of full-length genomes of types HPV16, 31, 35 and HPV 18, 45 and 59 in FASTA format. The HPV clusters are available: “HPV16_31_35_alignment.fa”; “HPV18_45_59_alignment.fa”. 2. A list of qPCR primers proposed is presented in Table 1. A graphic representation of amplification strategy is shown in Figure 1. Briefly, primers that produce overlapping amplicons less than 600 bp, that passed the secondary structure tests, designed with annealing temperatures above 58°C, were selected for experimental evaluation. Calibration curves were generated using SYBR Green chemistry and serial dilutions of a DNA standard as template for each genotype evaluated. For ORF HPV18 E7, the amplification plots, including the melt curve analysis are shown in Figure 2. Primer pairs producing multiple products (Figure 3), are not recommended. Complete information for primer pair used in real-time PCR assays are described in Table 2. All this information is presented in Figures and Tables folders.THIS DATASET IS ARCHIVED AT DANS/EASY, BUT NOT ACCESSIBLE HERE. TO VIEW A LIST OF FILES AND ACCESS THE FILES IN THIS DATASET CLICK ON THE DOI-LINK ABOV