Difference between revisions of "Part:BBa K2262001"
 
(2 intermediate revisions by the same user not shown)
Line 13: Line 13:
 
<h1>'''Introduction'''</h1>
 
<h1>'''Introduction'''</h1>
 
&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 
&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
−
This peptide is from the MiAMP family. It is an anti-fungal peptide which inhibits the growth of a variety of fungi and has no activity against E.coli. The mechanism of the peptide is to bind specifically to beta-(1,3)-glucans in fungal cell walls through conserved polar and aromatic residues. NCTU_Formosa used it to be one of a training data for creating our anti-fungal peptide scoring card.  
+
This peptide derives from the MiAMP family. It is an anti-fungal peptide which inhibits the growth of a variety of fungi and shows no antimicrobial effects. The mechanism of the peptide is to bind specifically to beta-(1,3)-glucans in fungal cell walls through conserved polar and aromatic residues [1]. NCTU_Formosa used it to be one of a training data for forming the antifungal peptide scoring card.
  
 
<br>
 
<br>
−
 
  
 
<h1>'''Experiment'''</h1>
 
<h1>'''Experiment'''</h1>
 
&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 
&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
−
We put T7 promoter, RBS, and MiAMP 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+MiAMP1 is around 250 ~ 500 b.p.. Its PCR product is around 500 ~ 750 b.p..
+
We put T7 promoter, RBS, and MiAMP 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+MiAMP1 is 309 b.p. Its PCR product is 621 b.p.
  
 
<br>
 
<br>
  
 
[[File:Agarose gel MiAMP1.png|200px|thumb|center|'''Figure 2.''' P<sub>T7</sub> + RBS + MiAMP1 <br>
 
[[File:Agarose gel MiAMP1.png|200px|thumb|center|'''Figure 2.''' P<sub>T7</sub> + RBS + MiAMP1 <br>
−
The DNA sequence length of P<sub>T7</sub> + RBS + MiAMP1 is around 250 ~ 500 b.p.. The size of its PCR product should be close to 600 ~650 b.p..]]
+
The DNA sequence length of P<sub>T7</sub> + RBS + MiAMP1 is around 250 ~ 500 b.p. The size of its PCR product should be close to 600 ~650 b.p.]]
  
 
<br>
 
<br>
Line 32: Line 31:
 
<h1>'''Reference'''</h1>
 
<h1>'''Reference'''</h1>
  
−
McManus, A.M.; Nielsen, K.J.; Marcus, J.P.; Harrison, S.J.; Green, J.L.; Manners, J.M.; Craik, D.J. "MiAMP1, a novel protein from Macadamia integrifolia adopts a Greek key beta-barrel fold unique amongst plant antimicrobial proteins." J. Mol. Biol. 1999, 293, 629-638  
+
[1] McManus, A.M.; Nielsen, K.J.; Marcus, J.P.; Harrison, S.J.; Green, J.L.; Manners, J.M.; Craik, D.J. "MiAMP1, a novel protein from Macadamia integrifolia adopts a Greek key beta-barrel fold unique amongst plant antimicrobial proteins." J. Mol. Biol. 1999, 293, 629-638
 +
 
 +
 
  
  

Latest revision as of 15:07, 7 December 2017


T7 Promoter+RBS+MiAMP1


Figure 1. PT7+RBS+MiAMP1+terminator



Introduction

      This peptide derives from the MiAMP family. It is an anti-fungal peptide which inhibits the growth of a variety of fungi and shows no antimicrobial effects. The mechanism of the peptide is to bind specifically to beta-(1,3)-glucans in fungal cell walls through conserved polar and aromatic residues [1]. NCTU_Formosa used it to be one of a training data for forming the antifungal peptide scoring card.


Experiment

      We put T7 promoter, RBS, and MiAMP 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+MiAMP1 is 309 b.p. Its PCR product is 621 b.p.


Figure 2. PT7 + RBS + MiAMP1
The DNA sequence length of PT7 + RBS + MiAMP1 is around 250 ~ 500 b.p. The size of its PCR product should be close to 600 ~650 b.p.



Reference

[1] McManus, A.M.; Nielsen, K.J.; Marcus, J.P.; Harrison, S.J.; Green, J.L.; Manners, J.M.; Craik, D.J. "MiAMP1, a novel protein from Macadamia integrifolia adopts a Greek key beta-barrel fold unique amongst plant antimicrobial proteins." J. Mol. Biol. 1999, 293, 629-638



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]