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In Situ Cell-Free Expression of Unknown Proteins: Gateway Technology for Protein Cloning and Testing

Explore the potential of in situ cell-free expression for unknown proteins using Gateway Technology. Move genes effortlessly via recombination methods. Test proteins in varied ways, from assays in bacteria to analyzing interactions and functions in mammalian cells. Use Xho1 & BamHI digestion for verification. Detect in situ cell-free protein expression with WB Bead Suspension Arrays, coupling anti-GST antibodies to beads and capturing GST-tagged proteins. Advance your research with efficient protein production and testing capabilities.

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In Situ Cell-Free Expression of Unknown Proteins: Gateway Technology for Protein Cloning and Testing

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  1. In situ cell-free expression of unknownproteins Progres Meeting Madrid, 2012

  2. 16 Genes onChromosome 16 16 More than85 % of proteincoding genes of chromosome 16 are expressed in lymphoidcells, epitelial cells and fibroblasts.

  3. 16 Genes onChromosome 16 16 More than85 % of proteincoding genes of chromosome 16 are expressed in lymphoidcells, epitelial cells and fibroblasts.

  4. Gateway Technologyforcloning & subcloning Moving Genes by Recombination Master Only the Desired Product Survives Mix two plasmids and enzyme

  5. Testing Proteins Many Different Ways • Master Library  Many libraries • Any vector can be modified to accept genes

  6. Testing Single Proteins in Many Different Ways Perform assays using protein made in bacteria. What other proteins does this protein interact with? Where is the protein located in the cell? Test the protein’s function in mammalian cells. Produce lots of protein for experiments. Turn the protein on and off in mammalian cells.

  7. InsertVerificationby Xho1 & BamHIDigestion TMEM170 Marcador 1Kb WFDC1 MC1R LRRC29 KCNG4 IGSF6 CKLFSF2 GSG1L

  8. Detection of in situ cell-free proteinexpression WB BeadsSuspensionArrays (BBAs)

  9. Y Y Y Y Y Y Y Y Y Y Y Y Y Y Y Y Y Y Y Y Y Y Y Y BeadsSuspensionArrays Coupling of anti-GST antibody to beads Anti-GST Anti-mouse Mix Capture of GST- Detection re-aliquot tagged protein onto anti-GST beads Cell-free expression of GST-tagged DNA

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