Part:BBa_K4171011

PlacI-cysE*

Background

P_lacI-cysE* is constructed to express CysE* (BBa_K4171004), an enzyme involved in cysteine synthesis pathway that catalyzes serine into O-acytel-serine (OAS).

Usage

P_lacI-cysK-cysE* (BBa_K4171012) was cloned to convert OAS and selenide into selenocysteine, for the similarities shared between sulfide and selenide. Therefore, we first cloned P_lacI-cysE* (BBa_K4171011) and sent it to E. coli △cysE mutant with dysfunctional CysE, to confirm the function of CysE*.

Fig. 2. In vivo Sec synthesis pathway

Characterization

Fig. 2. Confirmation of P_lacI-cysE* (BBa_K4171011) by colony PCR. M: Marker; Lane 1: P_lacI-cysE* (1263bp).

To evaluate CysE* function, we transformed BBa_K4171011 into E. coli △cysE mutant and observed its growth curve. Theoretically, once we conducted transformation successfully, the plasmid should compensate for the mutant, which is corresponding to the result shown below.

Fig. 3. Confirmation of CysE* function by transforming P_lacI-cysE* (BBa_K4171011) into E. coli △cysE mutant.

Sequence and Features

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 AgeI site found at 918
  
• 1000
  COMPATIBLE WITH RFC[1000]