We investigated the design of potential immunogenic peptides of the gSG6 protein using bio-informatic tools. The strategy was i) to identify potential immunogenic epitopes predicted by algorithms; and ii) to research the specificity of An. gambiae gSG6 peptide sequence compared to the genome/ Expressed Sequence Tag (EST) libraries of other organisms.
This analysis was based on the amino acid sequence of mature An. gambiae gSG6 (“UniProtKB/TrEMBL:Q9BIH5” and “gi:13537666”, [23] (link)).
The identification of putative linear B-cell epitopes of An. gambiae gSG6 was performed by computerized predictions of antigenicity based on physico-chemical properties of the amino-acid sequences with the BcePred database [24] and with the FIMM database [25] (link). We also identified the MHC class 2 binding regions using the ProPred-2 online service [26] (link).
Sequence alignments were done with the Tblastn program in Vectorbase database [27] (link) which enabled comparing a sequence of gSG6 peptides with known genomes or EST libraries of Aedes aegypti, Ixodes scapularis, Culex pipiens, Pediculus humanus, Glossina morsitans, Rhodnius prolixus, Lutzomia longipalpis and Phlebotomus papatasi. Concomitantly, we investigated sequence alignments with the Blast program to compare the gSG6 peptides sequence with all non-redundant GenBank CDS database [28] (link).
Peptides were synthesized and purified (>80%) with Genosys (Sigma-Genosys, Cambridge, UK) with an added N-terminal biotin. All peptides were shipped lyophilized and they were resuspended in 0.22 µm filtered milliQ water and stored in aliquots at −80°C.
This analysis was based on the amino acid sequence of mature An. gambiae gSG6 (“UniProtKB/TrEMBL:Q9BIH5” and “gi:13537666”, [23] (link)).
The identification of putative linear B-cell epitopes of An. gambiae gSG6 was performed by computerized predictions of antigenicity based on physico-chemical properties of the amino-acid sequences with the BcePred database [24] and with the FIMM database [25] (link). We also identified the MHC class 2 binding regions using the ProPred-2 online service [26] (link).
Sequence alignments were done with the Tblastn program in Vectorbase database [27] (link) which enabled comparing a sequence of gSG6 peptides with known genomes or EST libraries of Aedes aegypti, Ixodes scapularis, Culex pipiens, Pediculus humanus, Glossina morsitans, Rhodnius prolixus, Lutzomia longipalpis and Phlebotomus papatasi. Concomitantly, we investigated sequence alignments with the Blast program to compare the gSG6 peptides sequence with all non-redundant GenBank CDS database [28] (link).
Peptides were synthesized and purified (>80%) with Genosys (Sigma-Genosys, Cambridge, UK) with an added N-terminal biotin. All peptides were shipped lyophilized and they were resuspended in 0.22 µm filtered milliQ water and stored in aliquots at −80°C.
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