Part:BBa_K2309028

3xAMPs with LL-37 plus his tag in Lactococcus Latics (optimized)

Our design using the tandem repeats strategy to express several copies of each anti-microbial peptide (LL-37, GF-17, and Grammistin-Pp1). Two gene will recombinant together in tandem repeats. Replication slippage could be formed between homology sequence between long space (50-100bp in general). In our project, we put repeat sequences together as ‘---AAA---BBB---CCC---‘ rather than ‘---ABC---ABC---ABC---‘ strategy. Therefore, recombination could happened between homology sequence. The replication slippage will not formed (Gemayel. et al, 2010).

 Why adding histidine-tag on LL-37? We added a 6x His tag at the end of LL-37 and cloned the three copies of the tagged LL-37 gene at the end of the whole construct as shown in Figure 5 below. This design is used for the detection of the expression of the peptides from the whole construct. It is very likely that if the peptide LL-37 plus the histidine tag was detected by immunoblotting, that the other peptides located upstream of LL-37 (GF-17, Grammistin-Pp1, and LL-37) were also translated.

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