Part:BBa_K542010

Enhanced Lumazine Synthase (ELS)

The part was synthesized by [http://www.biobasic.com/ Bio Basic Inc.] into the pET28a plasmid vector with an N-terminal His-tag. The Enhanced Lumazine Synthase (ELS) without the His-tag was moved into the pSB1C3 plasmid vector by the Lethbridge 2011 team.

The size of the cavity of the Lumazine Synthase microcompartment was enlarged via directed evolution (Wörsdörfer et al., 2011). The compartment formed by this polypeptide has a larger cavity than the Lumazine Synthase microcompartment submitted by the Lethbridge 2009 team (BBa_K249002). A larger interior allows for the option of localizing larger and/or more proteins into the cavity of the compartment.

The His-tagged ELS in pET28a was transformed into E. coli BL21 (DE3) and overexpressed.

Figure 1: 12.5% SDS-PAGE of His-tagged ELS in BL21 (DE3) Overexpression. Samples were taken at 1 hour increments after IPTG induction. Lane 1: zero hours; Lane 2: 1 hour; Lane 3: 2 hours; Lane 4: 3 hours. Clear overexpression observed near ~25.0 kD marker.

Reference:
Wörsdörfer, B., Woycechowsky, K.J., and Hilvert, D., (2011). Directed Evolution of a Protein Container. Science. 331: 589-592.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 5
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]