Difference between revisions of "Part:BBa K2262011"
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__NOTOC__ | __NOTOC__ | ||
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<h1>'''Introduction'''</h1> | <h1>'''Introduction'''</h1> | ||
| − | It has been known as | + | 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. |
<h1>'''Experiment'''</h1> | <h1>'''Experiment'''</h1> | ||
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<p style="padding:1px;font-size:16px"><b>1. Fungal Test </b></p> | <p style="padding:1px;font-size:16px"><b>1. Fungal Test </b></p> | ||
<p style="padding:1px;font-size:14px"><b>(1) Inhibition Zone </b></p> | <p style="padding:1px;font-size:14px"><b>(1) Inhibition Zone </b></p> | ||
| + | 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. | ||
| + | [[File:Inhibition zone B-1E.png|800px|thumb|center|'''Figure 2.''' The result shows that B-1E will affect the growth of mycelium. ]] | ||
<p style="padding:1px;font-size:14px"><b>(2) Spore Germination </b></p> | <p style="padding:1px;font-size:14px"><b>(2) Spore Germination </b></p> | ||
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<p style="padding:1px;font-size:14px"><b>(3) Botany Experiment </b></p> | <p style="padding:1px;font-size:14px"><b>(3) Botany Experiment </b></p> | ||
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<p style="padding:1px;font-size:16px"><b>2. Cloning </b></p> | <p style="padding:1px;font-size:16px"><b>2. Cloning </b></p> | ||
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<span class='h3bb'>Sequence and Features</span> | <span class='h3bb'>Sequence and Features</span> | ||
<partinfo>BBa_K2262011 SequenceAndFeatures</partinfo> | <partinfo>BBa_K2262011 SequenceAndFeatures</partinfo> | ||
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Revision as of 15:48, 30 October 2017
T7 Promoter+RBS+B-1E
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.
(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
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]