Difference between revisions of "Part:BBa K2599008"
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===Experiment Result===
  
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The amino acid was determined to be as follows:X-Gly-Leu-Gly-Leu-Trp-Gly-Asn-Lys-Gly-Cys-Ala-Thr-Cys-Ser-(sequence; see text) Ile-Gly-Ala-Ala-Cys-Leu-Val-Asp-Gly-Pro-Ile-Pro-Asp-Glx-Ile-Ala-Gly-Ala. The analyses of cross-linking structures revealed that there were linkages between the amino- and carboxyl-termini and between the Cys-19 and the Glx-28 residues through an unknown residue with a residue weight of 163. Consequently, subtilosin A was deduced to be a cyclic peptide antibiotic with a novel cross-linking structure.
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<b>Cloning</b>
  
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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.
  
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Figure 2 structure picture
 
  
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Figure 2 PCR
  
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===Experiment Result===
 
  
  

Revision as of 06:56, 18 September 2018


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


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1055
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 778
    Illegal AgeI site found at 868
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 698