Team:Washington/Accomplishments

From 2009.igem.org

(Difference between revisions)
Line 83: Line 83:
*BBa_K215210 Display system (1 trans-membrane) in protein expression vector
*BBa_K215210 Display system (1 trans-membrane) in protein expression vector
*BBa_K215211 Display system (5 trans-membrane) in protein expression vector
*BBa_K215211 Display system (5 trans-membrane) in protein expression vector
-
*BBa_K215260
+
*BBa_K215260 BBa_K215210 fused to Streptavidin
-
*BBa_K215261
+
*BBa_K215261 BBa_K215211 fused to Streptavidin
Current Status: In progress
Current Status: In progress

Revision as of 04:49, 8 October 2009

Uw title logo.png


Target structure.png

BBa_K215002 Is a BioBrick part that was made to work in conjunction with the Secretion System (BBa_K215107). When used it creates a fusion protein, that contains the prtB secretion signal recognized by the secretion system. It also fuses to the protein a nano-tag that is able to bind Streptavidin, TEV protease site (for isolating your protein from the fusion) and Histidine Epitote Tags for easy protein purification.
This part is composed of:

BBa_K215000

  • Protein Expression Vector

BBa_K215001

  • Fusion protein sequence


This part is used in: BBa_K215010

  • GFP fusion protein

BBa_K215011

  • BBa_K215090 (organic phosphotriesterase) fusion protein

BBa_K215091

Current Status: Complete

Future directions:

  • Make more universal, to work with all proteins
  • Optimize secretion-tag and nano-tag for fusion proteins
Secretion structure.png

BBa_K215107 Is a BioBrick version of the Type 1 secretion system of Erwinia Chrysanthemi. It secretes proteins that contain the prtB C-terminus Epitote tag. To create a fusion protein with the prtB tag see BBa_K215002.

This part is composed of: BBa_K215100

  • Gene encoding prtD

BBa_K215101

  • Gene encoding prtE

BBa_K215102

  • Gene encoding prtF

BBa_K215103

  • BBa_K21500 + BBa_K21501 prtD + prtE

BBa_K215104

  • BBa_K25103 + BBa_K25102 prtD + prtE + prtF

BBa_K215105

  • BBa_K215104 + BBa_B0014 in PSB1AK3 prtDEF + Terminator

BBa_K215106

  • BBa_K215104 + BBa_B0014 in PSB3T5 prtDEF + Terminator

BBa_K215108

  • BBa_J23114 + BBa_K215106 Medium Promoter + prtDEF-Terminator

BBa_K215109

  • BBa_J23113 + BBa_K215106 Weak Promoter + prtDEF-Terminator

Current Status: In progress

Future directions:

- Optimize RBS's

- Move secretion sequence into new plasmid, not TetR

- Try variety of cell lines.

Display structure.png

BBa_K215200 is a Display vector that will allow for the presentation of any protein onto the surface of a cell by fusing the protein to Outer Membrane Protein A (OMPA). The fusion protein contains a GS peptide linker to space the displayed protein away from the outer membrane and also contains a TEV protease site to allow for purification of the displayed protein.

This Part is composed of:

  • BBa_K215201 Display Vector with 5 trans-membrane OMPA
  • BBa_K215250 Coding Sequence for fusion protein scaffold, 1 trans-membrane OMPA
  • BBa_K215251 Coding Sequence for fusion protein scaffold, 5 trans-membrane OMPA


This Part is used in:

  • BBa_K215210 Display system (1 trans-membrane) in protein expression vector
  • BBa_K215211 Display system (5 trans-membrane) in protein expression vector
  • BBa_K215260 BBa_K215210 fused to Streptavidin
  • BBa_K215261 BBa_K215211 fused to Streptavidin

Current Status: In progress

Future Directions:

- Use new CDS display vector to display streptavidin

- Design monomeric protein to bind peptide sequence

- Utilize other proteins on CDS to test for activity of other proteins

Biobricks Submitted Current Status Report Future Directions


Quick overview of what was done and the status of everyting. Maybe 3 paragraphs (display, secretion, conclusions)