Composite

Part:BBa_K2262011

Designed by: CHEN, PENG-TZU   Group: iGEM17_NCTU_Formosa   (2017-10-24)
Revision as of 15:49, 30 October 2017 by Peng-Tzu (Talk | contribs)


T7 Promoter+RBS+B-1E

Figure 1. PT7+RBS+B-1E+terminator

Introduction

It has been known as an antibacterial peptide that has activity against representative Gram-negative and Gram-positive bacterial species. Its grade given by scoring card ,which was created by NCTU_Formosa, shows it may also have an anti-fungal function.

Experiment

1. Fungal Test


(1) Inhibition Zone

We put some mycelium on a Potato dextrose agar plate, PDA plate, and cultivated it at the point of 20℃ for culturing Botrytis cinerea, the fungi we used, for 2 days. Ensuring the mycelial colony has developed, we put the sterile blank paper which the different concentrations of B-1E and HEPES, as a negative control, were added respectively. The place where peptide been but will appear a hole after 12 ~ 18 hours if the peptide will affect the growth of mycelium. The result shows that the place we put B-1E will have a hole. It means the peptide will affect the growth of mycelium.

Figure 2. The result shows that B-1E will affect the growth of mycelium.

(2) Spore Germination


(3) Botany Experiment


2. Cloning

We put T7 promoter, RBS, and B-1E together with pSB1C3 as a backbone. Then we conducted Taq PCR to check the size of the DNA sequence was right. The length of T7 promoter+RBS+Sa-AFP2 is around 100 ~ 200 b.p.. Its PCR product is around 250 ~ 500 b.p..


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]
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Categories
Parameters
None