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Part:BBa_K2278022
Lecrocin I antimicrobial peptide with Alpha-Factor Secretion Signal
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal XhoI site found at 244
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Introduction
This DNA biobrick was designed in order to produce in strain.
1- Biological background
Antimicrobial peptides are phygenitically ancient components of innate defense mechanisms of both invertebrates and vertebrates. In the context of growing prevalence of antibiotic-resistance of bacterial strain, the AMP can be considered as potential new therapeutical candidates. Leucrocin I from Siamese crocodile white blood cells shows a good antibacterial activity towards Vibrio cholerae. The peptide is a 7 amino acid residue : NGVQPKY with a molecular mass around 806.99 Da. The mechanism of action of the Leucrocin I has been observed with fluorescence and electron microscopy This cationic molecules and can target bacterium membranes, to create pores in it, leading to the lysis of the cells. The part was designed to constitutively produce the leucrocin I AMP with a yeast promoter. The α-factor sequence contains a RBS and a signal sequence to secrete the produced peptides. The pGAP promoter is used because it makes genome recombination easier in Pichia pastoris genome. The restriction enzyme sites are set up to extract individually each components of the plasmid. It belongs to the respond module in the Croc’n cholera project of iGEM INSA-UPS-France 2017 Mécanisme
2- Usage in iGEM projects
The BBa_K2278022 cames from the module of the Croc’n cholera project (team INSA-UPS-France 2017) It was designed to produce
The part includes
Experiments
1- Molecular biology
The gene was placed in silico under the control of the p promoter (BBa_R), a strong RBS (BBa_B0034) and a terminator (BBa_B1006). IDT performed the DNA synthesis and delivered the part as gBlock. The construct was cloned by conventional ligation into pSB1C3 plasmid and transformed into E. coli Dh5 alpha strain. X transformants were obtained.
Analysis of the restriction mapSequencing
2- Expression in vivo
sous titre
Protocole
Characterization
1- Validation of
descriptionmanip1
Image styléeInterprétation
manip2
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Discussion :
2. 2ème approche
brillante analyse
Discussion :
des perspectives éclectiques
BBa_K2278022None |