Difference between revisions of "Part:BBa K2686001:Experience"
 
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The part was designed and constructed in a pET14 vector. The sequence was confirmed using sanger sequencing. The part was expressed in the pET14 vector using a BL21 DE3 TX-TL (Sun et al., 2013) cell free expression system. After expression and purification, the part was validated in multiple ways. First, multiple SDS-PAGE experiments indicate that our expressed protein is at the correct hypothetical size, which was further confirmed by lysine BODIPY tRNA expression (32.96kDa for the monomer and 1.98MDa for the 60-mer). DLS measurements also show that our protein cage self-assembles into the 60-mer.
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The part was designed and constructed in the original pET14 vector from <bbpart>K2686001</bbpart> (Cassidy Armstutz et al., 2016, Addgene plasmid #86405). The sequence was confirmed using Sanger sequencing. The part was expressed in the pET14 vector using a BL21 DE3 TX-TL (Sun et al., 2013) cell free expression system. First, multiple SDS-PAGE experiments indicate that our expressed protein is at the correct hypothetical size, which was further confirmed by lysine BODIPY tRNA expression (32.16calckDa for the monomer and 1.93MDa for the 60-mer).
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===Applications of BBa_K2686001===
 
===Applications of BBa_K2686001===
  
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===References===
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Cassidy-Amstutz, C., Oltrogge, L., Going, C., Lee, A., Teng, P., Quintanilla, D., East-Seletsky, A., Williams, E. and Savage, D. (2016). Identification of a Minimal Peptide Tag for in Vivo and in Vitro Loading of Encapsulin. Biochemistry, 55(24), pp.3461-3468.

Latest revision as of 22:31, 17 October 2018


The part was designed and constructed in the original pET14 vector from K2686001 (Cassidy Armstutz et al., 2016, Addgene plasmid #86405). The sequence was confirmed using Sanger sequencing. The part was expressed in the pET14 vector using a BL21 DE3 TX-TL (Sun et al., 2013) cell free expression system. First, multiple SDS-PAGE experiments indicate that our expressed protein is at the correct hypothetical size, which was further confirmed by lysine BODIPY tRNA expression (32.16calckDa for the monomer and 1.93MDa for the 60-mer).

Applications of BBa_K2686001

User Reviews

UNIQ68aa78e7b0e756f9-partinfo-00000001-QINU UNIQ68aa78e7b0e756f9-partinfo-00000002-QINU

References

Cassidy-Amstutz, C., Oltrogge, L., Going, C., Lee, A., Teng, P., Quintanilla, D., East-Seletsky, A., Williams, E. and Savage, D. (2016). Identification of a Minimal Peptide Tag for in Vivo and in Vitro Loading of Encapsulin. Biochemistry, 55(24), pp.3461-3468.