TNT® Eukarayotic Cell-free Protein Expression Systems - Promega

CELL-FREE PROTEIN EXPRESSION
TnT ® in vitro Transcription/Translation Systems
References #4
Cell-Free Protein Expression: Protein Arrays
3. Rolf, A. et al. (2008) Proc. Natl. Acad. Sci. 105,
4364-69. In this reference they report the assembly
of a complete ORF collection (3887) for V. cholerae.
15% of this collection was transferred into relevant
GST expression vectors and arrayed as plasmid DNA
and subsequently expressed using Promega’s TnT
system. The resulting proteins were screened to
determine if they triggered a Toll-like recepetors
(TLR) using A549 cells containing a luciferase gene
transcribed under control of a NF-κβ response
promoter. Using this approach a novel TLR5 agonist
(FlaC) was characterized.
4. Motor, W. et al. (2009) Infection Immun. 77, 4877-
86. A comprehensive study of 266 gene products
of the 5570 ORFs of Psuedmonas aeruginosa that
were likely to reside in the outer membrane or
secreted into the surrounding media were screened
(using the NAPPA technology) for their antigenicity.
From this study it was possible to identify 12
proteins that trigger an adaptive immune response
in cystic fibrosis and acutely infected patients.
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