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| − | {{Aix-Marseille|title=Phagemids|toc=__TOC__}}
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| − | [[File:T--Aix-Marseille--pbluescript-SN.jpeg|400px|left]]
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| − | [[File:T--Aix-Marseille--pSB1C3-SN.jpeg|400px|centre]]
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| − | To deliver our toxin, either we created a phagemid that contain the oriM13 (BBa_K1445000) which will gives it the opportunity to be used in the phage construction, or we used the phagemid pBluescript II KS(+).
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| − | Both of those phagemid contain a M13 origin of replication and a gene for antibiotic resistance. We insert in both, a ''E.coli'' or ''X. fastidiosa'' promoter along with SuperNova gene.
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| − | ===''X. fastidiosa'' promoter===
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| − | [[File:T--Aix-Marseille--M13pIII-SoftBerry.jpeg|400px|right]]
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| − | Firstly, we found best bidirectional hit (BBH) between Escherichia coli str. K-12 substr. MG1655 genes and ''Xylella fastidiosa'' 9a5c ones. In order to have a strong constitutive promoter we look at highly expressed genes from ''E.coli''.<ref>S, K., J, M., A, C. & D, K. Characterizations of highly expressed genes of four fast-growing bacteria., Characterizations of Highly Expressed Genes of Four Fast-Growing Bacteria. J Bacteriol 183, 183, 5025, 5025–5040 (2001).</ref>
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| − | Secontly, with the tool rsat, for each gene selected we take the upstream sequence from the previous gene to the ATG And with the tool BPROM we choose the sequence with predicted box with the best score. We choose XF_RS01885 which is the BBH of purA, which code for an adenylosuccinate synthetase.
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| − | Finaly, we tried to find the ribosome binding site (RBS) consensus in ''Xylella fastidiosa''. To do so we search for the anti-Shine dalgarno sequence with ''Xylella fastidiosa'' 16S ribosomal RNA gene (accession number : NR_041779). The consus found is : AGGAGG. The RBS is supposed to be 6 to 12 nucleotide upstream the ATG. So we modified the sequence. And we added Rfc10 prefix and suffix region.
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