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Generation of Recombinant mCMVs

KG Kerstin M. Gergely
JP Jürgen Podlech
SB Sara Becker
KF Kirsten Freitag
SK Steffi Krauter
NB Nicole Büscher
RH Rafaela Holtappels
BP Bodo Plachter
MR Matthias J. Reddehase
NL Niels A. W. Lemmermann
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Recombinant viruses mCMV-SIINFEKL and mCMV-SIINFEKA were generated by two-step replacement BAC mutagenesis in the mCMV BAC plasmid pSM3frΔm157luc (48), replacing a sequence that codes for an endogenous Dd-presented antigenic peptide in the non-essential gene m164 with sequences coding for peptides SIINFEKL or SIINFEKA (49, 50). The recombinant mCMVs were reconstituted in MEF and were propagated for removal of BAC sequences and for amplification (51). Infectious virions were purified according to standard procedures (47). Reconstituted and purified virus derived from the parental BAC plasmid pSM3frΔm157luc served as a control virus, for the sake of brevity herein referred to as mCMV, despite features included for a multi-purpose usage not applying to this report.

Copyright and License information:  ©2021 Gergely, Podlech, Becker, Freitag, Krauter, Büscher, Holtappels, Plachter, Reddehase and Lemmermann
This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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