Part:BBa_K117002:Design

LsrA promoter (indirectly activated by AI-2)

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

Design Notes

LsrA is initially commercially synthesized with RBS BBa_B0034 by ATIbiotech in a blue script cloning vector. LsrA is then extracted using PCR with oligo fragments of DNA sequence 5'-GAATTATGTATACATAATGGCA-3' & 5'-GGTAAATATCCTTTAATTAAA-3' with Biobricks prefix and suffix respectively.

Source of Origin Sequence

Molecular cloning of a gene encoding the thermoactive levansucrase from Rahnella aquatilis and its growth phase-dependent expression in Escherichia coli Jeong-Woo Seo, Ki-Bang Song, Ki-Hyo Jang, Chul-Ho Kim, Bong-Hyun Jung, Sang-Ki Rhee * Microbial and Bioprocess Engineering Laboratory, Korea Research Institute of Bioscience and Biotechnology (KRIBB), PO Box 115, Yusong, Taejon 305 -600, South Korea

Pitfalls

Wild type K12 E coli contains identical LsrA gene sequence. Extraction of plasmids using Qiagen plasmid extraction kit produced high quality pure plasmids. However, PCR with designed oligo fragments were unable to produce the desired base pair length of DNA. Other attempts of PCR using Platinum PCR mix did not yield any results.

References

Molecular cloning of a gene encoding the thermoactive levansucrase from Rahnella aquatilis and its growth phase-dependent expression in Escherichia coli Jeong-Woo Seo, Ki-Bang Song, Ki-Hyo Jang, Chul-Ho Kim, Bong-Hyun Jung, Sang-Ki Rhee *Microbial and Bioprocess Engineering Laboratory, Korea Research Institute of Bioscience and Biotechnology (KRIBB), PO Box 115, Yusong, Taejon 305 -600, South Korea

Regulation of Uptake and Processing of the Quorum-Sensing Autoinducer AI-2 in Escherichia coli Karina B. Xavier and Bonnie L. Bassler* Department of Molecular Biology, Princeton University, Princeton, New Jersey