The analysis of the cDNA sequences showed no differences between the two races. The coding region of the Clpnl2 gene consisted of 1428 bp interrupted by four introns ranging in size from 60 to 87 bp (Figure 1). According to the 5′RACE analysis, a putative transcription starting point was localized [19], and the context of the start codon
ATG matched with the Kozak this website sequence for filamentous fungi [54]. Two possible regulatory sequences were identified in the 5′ untranslated region of Clpnl2: a putative regulatory sequence for binding to RAP1, which is a transcriptional factor that participates in the activation of transcription and the silencing of genes in yeast cells, located at position +54 [55] and a possible binding sequence for the transcription factor AbaA at position +69. AbaA binding sites have been observed in several genes that participate in the control of cell development in organisms such as A. nidulans and Tucidinostat order the dimorphic fungus P.
marneffei, where AbaA has been related to morphogenesis and dimorphism, respectively [56, 57]. These putative regulatory elements were localized downstream the transcription site which is an uncommon finding. Multiple binding sites to AbaA have been reported in cis regulatory regions and some downstream the transcription starting site in A. nidulans genes. No attempts were made in this study to determine the function of these elements. Due to the size of the promoter region of Clpnl2, it was not possible to locate more elements commonly found in genes encoding for pectinolytic enzymes. The 5′ and 3′ untranslated regions (5′UTR
and 3′UTR) were 129 and 563 bp, respectively. Two consensus sequences (AATAAA and TTTCACTGC) found in the terminal regions of eukaryotic mRNAs [58], and two of the three consensus sequences for yeast 3′-terminal regions (TAGT and YIT) [59] were detected in the Clpnl2 3′UTR. Figure 1 Nucleotide and deduced amino acid sequence of the Clpnl2 gene. Intron and exon sequences are in lowercase and uppercase, respectively. The signal peptide sequence is boxed. The possible Tangeritin binding sequences of RAP1 and AbaA are underlined with a dotted line. The putative transcription start point is underlined, and the putative Kozak sequence is shaded. The sequences of the 3′-terminal region are underlined. An asterisk (*) marks the translation stop codon. The potential N-glycosylation site is circled. This sequence has been deposited in the GenBank nucleotide sequence database under accession number JN034038. The Clpnl2 cDNA contains an ORF of 1140 nucleotides that https://www.selleckchem.com/products/verubecestat.html encodes a putative protein of 379 aa with a N-terminal secretion signal sequence of 19 amino acids, according to the SignalP 3.0 web server [41]. A protein of molecular mass 37.4 kDa and a pI of 9.1 was calculated, and one potential N-glycosylation site was located at position 110 (ExPASy Proteomics Server) [42].