Premium
Conformational analysis of a set of selectively screened apoptotic related genes in a SEB induced human PBMC module
Author(s) -
Freehauf Emily Ann,
Jett Marti,
Mendis Chanaka
Publication year - 2006
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.20.4.a119
Subject(s) - gene , proteases , peripheral blood mononuclear cell , gene expression , apoptosis , biology , caspase , microbiology and biotechnology , genetics , programmed cell death , enzyme , in vitro , biochemistry
Here we attempt to characterize a set of apoptotic related genes that are key to the pathogenesis of Staphylococcal Enterotoxin B (SEB) induced lethal shock in human peripheral blood mononuclear cells (PBMC). We have identified and verified a set of genes that are important in the humans and non‐human primates. Gene expression patterns of key proteases have also been confirmed using Elisa assays. By using a set of genes previously identified through differential display (DD) and micro‐arrays we attempted to carry out a conformational study of SEB induced apoptotic events in the following manner. First, we carried‐out an extensive literature search to characterize apoptosis related genes identified by DD and micro‐arrays. Second, we designed primers using stringent parameters, third we confirmed the gene expression pattern using RT‐PCR and finally we validated the gene expression patterns by further investigation of protein expression using ELISA assays. We were able to confirm the previously observed gene expression patterns of heparanase precursor, Caspase 3, Caspase 6, Caspase 7, Caspase 8, Ubiquitin specific protease and SOD genes through RT‐PCR, protein expression pattern of a set of caspases through ELISA assays and correlated the functions of genes to SEB induced apoptosis related events. We believe that our investigation may reveal crucial information to better understand the mechanisms of SEB induced apoptotic events. Funds for research were provided by the Pioneer Undergraduate Research Foundation.