Part:BBa_K2599008
T7 Promoter+RBS+Subtilosin+intein+CBD
NCTU_Formosa 2018 designed a composite part encoding the Subtilosin sequence (BBa_K2599000), and then combined with a T7 promoter (BBa_I712074), a ribosome binding site (BBa_B0034), intein and chintin binding domain (CBD). Further information of our peptide can be found on our design page.
Figure 1 biobrick picture
Introduction
Bacillus subtilis produces an anionic bacteriocin called Subtilosin that possesses antibacterial activity against certain gram-positive bacteria. The bacteriocins are a group of anitmicrobial peptides that are often distinguished from traditioinal antibiotics by their narrow range of avtivity against closely related bacteria.
Experiment Result
Cloning
We conbined our toxic gene to pSB1C3 backbone and conducted PCR to check the size of our part. The Subtilosin sequence length is around 147 b.p. For the composite part, the sequence length should be near at 1191 b.p.
Figure 2 PCR
Expressing
We chose E. coli 2566 strain to express our antimicrobial peptides.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 1055
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 778
Illegal AgeI site found at 868 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 698
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