Difference between revisions of "Part:BBa K2309028:Design"

 
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===Design Notes===
 
===Design Notes===
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Three copy of Anti-Microbial Peptide
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Our design (BBa_K2309028)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).
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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).
  
  
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 Why adding histidine-tag on LL-37?
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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 .
  
 
===Source===
 
===Source===
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===References===
 
===References===
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Gemayel, R., (2010) ‘Variable Tandem Repeats Accelerate Evolution of Coding and Regulatory Sequences’, Annual Review of Genetics, ResearchGate [Online]. DOI:  10.1146/annurev-genet-072610-155046

Latest revision as of 07:47, 1 November 2017


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


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

Our design (BBa_K2309028)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 .

Source

artificial design

References

Gemayel, R., (2010) ‘Variable Tandem Repeats Accelerate Evolution of Coding and Regulatory Sequences’, Annual Review of Genetics, ResearchGate [Online]. DOI: 10.1146/annurev-genet-072610-155046