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Series GSE49605 Query DataSets for GSE49605
Status Public on Oct 01, 2013
Title Screening for novel immungenic proteins of Salmonella Enteritidis 125109
Platform organism Escherichia coli
Sample organism Salmonella enterica subsp. enterica serovar Enteritidis
Experiment type Protein profiling by protein array
Summary The screening of a cDNA derived expression library of Salmonella Enteritidis 125109 expressed in E. coli using a fusion construct and specific HaloTag interaction to a modified surface is shown. Thus, 1536 different clones were screened including positive (fimA) and negative (argC, gapA) reference proteins. The goal of the screening was to identify potential novel immunogenic proteins from S. Enteritidis by selecting clones showing a high signal intensity in comparison to the known antigens used as positve markers. Afterwards, the most promising clones were sequenced to identify the gene and corresponding protein and these proteins were then investigated further. Consequently, 9 novel immunogenic proteins could be identified.
 
Overall design In total 1536 different lysates were spotted on different microarray slides. Each slides contained 3600 distinct spots, seperated into two compartments of 1800 spots each. Each compartment comprised quadruplicate replicates of each sample lysate with controls being analysed with more replicates. As controls we used: fimA from 3 different samples (40 replicates) as positive reference proteins, as it has been described as immunogenic before. GapA from Klebsiella pneumoniae and Campylobacter jejuni (40 replicates) as negative reference proteins, additionally two sets of E.coli cell lysates without fusionprotein expressed, namely Acella electrocompetent cells (40 replicates) and KRX (32 replicates). Last but not least a buffer control (24 replicates) was included. Therefore, each set of replicate slides contained 384 different samples. Each screening was performed with three replicate slides. Consequently, a total number of 12 slides were screened (4 sets of samples x 3 replicates each). For identification rabbit polyclonal antibody to S. enterica (Abcam ab35156) as primary and Goat polyclonal to Rabbit IgG conjugated with ChromeoTM-546 (Abcam ab60317) as secondary antibody were used. The top compartment was incubated first with primary antibody, while the bottom compartment was incubated with PBS at the same time. Afterwards both compartments were incubated with secondary antibody.
 
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Submission date Aug 06, 2013
Last update date Oct 02, 2013
Contact name Sebastian Hoppe
E-mail(s) sebastian.hoppe@izi-bb.fraunhofer.de
Organization name Fraunhofer Institute for Cell Therapy and Immunology
Department Bioanalytics and Biosensorics
Lab Molecular Bioengineering
Street address Am M├╝hlenberg 13
City Potsdam
ZIP/Postal code 14476
Country Germany
 
Platforms (1)
GPL17538 IBMT Salmonella Enteritidis library screening
Samples (12)
GSM1202499 immunoscreening S. Enteritidis 1 (415842)
GSM1202500 immunoscreening S. Enteritidis 1 (415843)
GSM1202501 immunoscreening S. Enteritidis 1 (4169664)
Relations
BioProject PRJNA214479

Download family Format
SOFT formatted family file(s) SOFTHelp
MINiML formatted family file(s) MINiMLHelp
Series Matrix File(s) TXTHelp

Supplementary file Size Download File type/resource
GSE49605_RAW.tar 1.5 Mb (http)(custom) TAR (of TXT)
Raw data provided as supplementary file
Processed data included within Sample table

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