Cloning of hCMV iE1 Proteins

The cDNAs encoding hCMV iE1^1–328 and iE1^1–289 fused to an N-terminal FLAG tag were obtained by gene synthesis (Integrated DNA Technologies, Inc.) and then cloned into the pCDNA.3 vector (Invitrogen) (pCDNA3-FLAG-hCMV iE1^1–382 [BUG 3779; Bacteria-Cell Interactions laboratory's bacteria collection] and pCDNA3-FLAG-hCMV iE1^1–289 [BUG 3780]). The plasmids encoding YFP-CBD, a yellow fluorescent protein (YFP) chimera protein of the cell wall binding domain (CBD) from the Listeria phage endolysin Ply118 (BUG 2305; kind gift from J. Swanson, University of Michigan Medical School, Ann Arbor, MI, USA), and Sp100 (pSG5-Sp100^WT [BUG 4134] and its derivative pSG5-Sp100^K297R [BUG 4135]) have been described previously (11 (link), 57 (link)).

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Ribet D., Lallemand-Breitenbach V., Ferhi O., Nahori M.A., Varet H., de Thé H, & Cossart P. (2017). Promyelocytic Leukemia Protein (PML) Controls Listeria monocytogenes Infection. mBio, 8(1), e02179-16.

Publication 2017

gene synthesis pCDNA.3 vector pCDNA3

Bacteria bacteria Cdnas Cell Cell interactions Cell wall Chimera Endolysin Hcmv Listeria Phage Plasmids Protein domain Sp100 Synthesis gene Vector

Corresponding Organization :

Other organizations : Institut Pasteur, Inserm, Université Paris Cité, Délégation Paris 7, Centre National de la Recherche Scientifique, Collège de France

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Variable analysis

independent variables

Expression of hCMV iE1^1–328 and iE1^1–289 fused to an N-terminal FLAG tag

dependent variables

Not explicitly mentioned

control variables

Expression of YFP-CBD, a yellow fluorescent protein (YFP) chimera protein of the cell wall binding domain (CBD) from the Listeria phage endolysin Ply118
Expression of Sp100^WT and Sp100^K297R

positive controls

YFP-CBD (BUG 2305)
Sp100^WT (pSG5-Sp100^WT [BUG 4134])

negative controls

Sp100^K297R (pSG5-Sp100^K297R [BUG 4135])

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