The signal peptide was situated about the N terminal of the two proteins. Six conservative cystein residues have been current during the positions 29,45,50,97,104 and 120 within the alignment. Comparison in the allergen like protein and tick ML domain containing protein with all the sequences from the related proteins through the family members uncovered that allergen like protein belongs rather to group II of your ML protein family members that is composed of Npc2, seven mite big allergen proteins, eight D. melanogaster proteins and 5 C. elegans proteins. The tick ML domain containing protein was assigned to group I that contains human MD 1 and MD two proteins and their orthologs. The function from the gut expressed ML proteins in tick is unknown, however it is obvious they is likely to be involved in host response to pathogen parts and mediate defensive reactions. Identification and molecular characterization of novel defensin gene.
the first annotation of two isoforms along with the presence of introns in genomic sequence of really hard tick Ixodes ricinus N. Rudenko, M. Golovchenko, a replacement L. Grubhoffer Faculty of Biological Sciences, University of South Bohemia and Division of Molecular Ecology of Parasite, Biological Center, Institute of Parasitology AS CR, Cesk? Budejovice, 37005, Czech Republic defensin gene, encoding the 8231 Da prepropeptide, 74 residues in total, which include signal peptide of 22 residues plus a propeptide of 15 amino acids, followed by a mature peptide of 37 residues, was isolated through the cDNA subtracted library of very hard tick Ixodes ricinus. Alignment of the mature area showed similarities to defensins from other species of tough ticks, ranging from 77% for I. scapularis to 56% to get a. hebraeum. Similarity to four described defensins from soft ticks O. moubata was 61 63% in a mature peptide.
The translated sequences of various recombinants through the exact same cDNA library indicated the presence of two isoforms a total noob of your I. ricinus defensin together with the approximate frequency of appearance as 4.one. The predominant form on the peptide is made up of glutamine at position 23, glutamic acid at position 25 and phenylalanine at position 45, substituted by glutamic acid and aspartic acid in the propeptide region, and arginine in the mature peptide inside the 2nd isoform. Regardless of whether these substitutions have an impact on the properties of peptide is currently unknown. I. ricinus defensin gene was strongly induced only in the midgut following infection with Borrelia burgdorferi. Defensin cDNA was noticed to become 225 bp, on the basis of which the primers for genomic PCR were created. Analysis of 926 bp of genomic sequence showed that I. ricinus defensin PS-341 gene calls for three exons, which are separated by two introns.