Difference between revisions of "Part:BBa K2804001:Design"
(→‎Design Notes)
 
(8 intermediate revisions by the same user not shown)
Line 1: Line 1:
 
__NOTOC__
 
__NOTOC__
−
<partinfo>BBa_K2804002 short</partinfo>
+
<partinfo>BBa_K2804001 short</partinfo>
  
−
<partinfo>BBa_K2804002 SequenceAndFeatures</partinfo>
+
<partinfo>BBa_K2804001 SequenceAndFeatures</partinfo>
  
  
 
===Design Notes===
 
===Design Notes===
−
Because the N-terminal domain of BMP2 has been mostly fused to proteins, we have decided to fuse it N-terminally to the CBD cipA.
+
BMP2 was fused to the C-terminus of CBD cipA because the osteogenic activity of the C-terminal domain of BMP2.
 +
 
 +
It was necessary to add basepairs at the N-terminus and C-terminus of the two gBlocks (CBD cipA under the control of LacI promoter and BMP2) in order to allow the cleavage by EcoRI and PstI. Then, a fusion assembly was done using the psb1c3 as cloning vectorof the fusion CBD cipA-BMP2.
  
 
===Source===
 
===Source===
  
−
Synthetized from IDT.
+
Fusion assembly.
  
 
===References===
 
===References===
−
Schmoekel, H.G., Weber, F.E., Schense, J.C., Grätz, K.W., Schawalder, P. & Hubbell, J.A. (2005). Bone repair with a form of BMP-2 engineered for incorporation into fibrin cell ingrowth matrices. Biotechnology and Bioengineering; 89(3):253-62.
+
Schmoekel, H.G., Weber, F.E., Schense, J.C., Grätz, K.W., Schawalder, P. & Hubbell, J.A. (2005). Bone repair with a form of BMP-2 engineered for incorporation into fibrin cell ingrowth matrices. Biotechnology and Bioengineering. 5;89(3):253-62.
 +
New England Biolabs. Cleavage Close to the End of DNA Fragments (oligonucleotides). https://www.neb.com/-/media/nebus/files/chart-image/cleavage_olignucleotides_old.pdf?la=en

Latest revision as of 02:21, 15 October 2018

CBD cipA fused to BMP2 under the control of LacI promoter


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 230
    Illegal AgeI site found at 1106
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

BMP2 was fused to the C-terminus of CBD cipA because the osteogenic activity of the C-terminal domain of BMP2.

It was necessary to add basepairs at the N-terminus and C-terminus of the two gBlocks (CBD cipA under the control of LacI promoter and BMP2) in order to allow the cleavage by EcoRI and PstI. Then, a fusion assembly was done using the psb1c3 as cloning vectorof the fusion CBD cipA-BMP2.

Source

Fusion assembly.

References

Schmoekel, H.G., Weber, F.E., Schense, J.C., Grätz, K.W., Schawalder, P. & Hubbell, J.A. (2005). Bone repair with a form of BMP-2 engineered for incorporation into fibrin cell ingrowth matrices. Biotechnology and Bioengineering. 5;89(3):253-62. New England Biolabs. Cleavage Close to the End of DNA Fragments (oligonucleotides). https://www.neb.com/-/media/nebus/files/chart-image/cleavage_olignucleotides_old.pdf?la=en