Part:BBa_K4012018

RtTAL

RtTAL is a sequence codes for tyrosine ammonia-lyase or Phenylalanine, which catalyzes the non-oxidative deamination of L-phenylalanine and L-tyrosine to form trans-cinnamic acid and p-coumaric acid respectively with similar efficiencies, facilitating the commitment step in phenylpropanoid pathways that produce lignins, coumarins and flavonoids. Tyrosine ammonia-lyase is a lyase involved in Phenylpropanoid metabolism. The catalytic activity: L-tyrosine = (E)-4-coumarate + NH4+

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 2021
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

RtTAL in Level1 plasmid assembly

Figure 2: [1] A: the construction strategy; B: results of colony PCR; C: results of sequencing analysis of coding sequence RtTAL

he construction schematic of RtTAL sequence demonstrated in Fig.2. The initiation of the RtTAL sequence is done by promoter pRET2, with termination done by tSSA1. The sequence ConLS and ConR1 are connector sequences within the Level 1 plasmid assembly. Furthermore, type9-KVF and type9-VR are imposed to enact selection through colonies PCR, results shown in Fig.2B. The band length 3432bp match with expectations. The sequence analysis results show no significant mutations or deletions, representing the success of the assembly.

GbCHS in Level2 plasmid assembly

Figure 3: [2] A: The results of colony PCR of upstream transformant; B: The results of colony PCR of downstream transformant; C: Construction of catechin metabolic pathway; D：Construction of naringenin metabolic pathway

RtTAL is successfully inserted into the vector type9 C-mTurquoise-Ura.