Heterologous Expression of Tagged Proteins

For the expression of FLAG-tagged CpoS proteins, the tag was inserted between the KpnI and SalI sites of p2TK2-SW2 (59 (link)). DNA fragments coding for CpoS proteins and their promoters were amplified (Table S3) or obtained as synthetic gene blocks (IDT) and then inserted between the AgeI and KpnI sites of the vector. For the expression of MYC-tagged Incs, the Inc genes and their promoters were amplified (Table S3) and then inserted between the AgeI and NheI sites of vector p2TK2-SW2 (59 (link)). Plasmids enabling the expression of mCherry (59 (link)) or inducible expression of GFP11-tagged IncB (CT232) (58 (link)) were gifts from Isabelle Derré and Kevin Hybiske, respectively. Bacteria were transformed as described above. Expression of GFP11-tagged IncB was induced by the addition of anhydrotetracycline (Clontech).

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Meier K., Jachmann L.H., Türköz G., Babu Sait M.R., Pérez L., Kepp O., Valdivia R.H., Kroemer G, & Sixt B.S. (2023). The Chlamydia effector CpoS modulates the inclusion microenvironment and restricts the interferon response by acting on Rab35. mBio, 14(4), e03190-22.

Publication 2023

anhydrotetracycline

Anhydrotetracycline Bacteria Genes Gifts Plasmids Proteins Synthetic gene Vector

Corresponding Organization :

Other organizations : Umeå University, Centre de Recherche des Cordeliers, Université Paris Cité, Inserm, Institut Universitaire de France, Sorbonne Université, Institut Gustave Roussy, Duke University, Hôpital Européen, Hôpital Européen Georges-Pompidou, Assistance Publique – Hôpitaux de Paris

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Variable analysis

independent variables

FLAG-tagged CpoS proteins
MYC-tagged Incs
Expression of mCherry
Inducible expression of GFP11-tagged IncB (CT232)

dependent variables

Not explicitly mentioned

control variables

Not explicitly mentioned

controls

Positive control: Plasmids enabling the expression of mCherry (59 (link))
Positive control: Plasmids enabling the inducible expression of GFP11-tagged IncB (CT232) (58 (link))

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