| Products/Services Used | Details | Operation |
|---|---|---|
| Codon Optimization | Four genes inclusive of ipaB, ipaD, icsA, and stxB are origin ofIpaB, IpaD, IcsA, and StxB protein respectively. The first three selected for their attaching and expanding function besides B subunit of Stx which is involved in the invasion as the main toxin. Associated DNA and protein sequences acquired from online databases, GenBank and UniProt (Universal Protein Resource) separately in FASTA format, and EBI database recruited for multiple sequence alignments by its clustal omega tool. The four sequence were attached to each other by an appropriate repetition of EAAAK as a rigid linker. Theoretical analysis of DNA parameter optimisation was done by using Rare Codon Calculator tool of GenScript web page (http://www.genscript.com/cgi-bin/tools/ rare_codon_analysis) besides OPTIMIZER tool of HEG database (Puigbo et al., 2007, 2008). The synthetic sequence was designed for cloning and expression in E. coli according to the k12 table of codon usage database (Nakamura et al., 2000). Also, WebDSV translates tool recruited parallel to all above for manipulation of sequences (http://www.molbiotools.com/WebDSV/). | Get A Quote |
Shigellosis is a high burden gastrointestinal disease with an increased frequency of antibiotic resistance. Type III secretion apparatus (T3SA) are conserved among different species of Shigella; and IpaD, IpaB, and IcsA proteins participate in its function. Studies indicate shiga toxin as a virulence factor has a fundamental role in hemorrhagic colitis. Bioinformatics tools were recruited for aiding this purpose. In the level of the nucleosome, sequences choosing and optimising and in the phase of the transcriptome, some prediction in associate with mRNA form, also in step of the proteome, physicochemical parameter, best stability, first to third structures and model validation were some prediction performed in... More
