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Optimization of a Lambda-RED Recombination Method for Rapid Gene Deletion in Human Cytomegalovirus

National Center for Microbiology, Instituto de Salud Carlos III Majadahonda, 28220 Madrid, Spain
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Academic Editors: Joaquin Arino and Paola Brun
Int. J. Mol. Sci. 2021, 22(19), 10558; https://doi.org/10.3390/ijms221910558
Received: 20 August 2021 / Revised: 23 September 2021 / Accepted: 27 September 2021 / Published: 29 September 2021
Human cytomegalovirus (HCMV) continues to be a major cause of morbidity in transplant patients and newborns. However, the functions of many of the more than 282 genes encoded in the HCMV genome remain unknown. The development of bacterial artificial chromosome (BAC) technology contributes to the genetic manipulation of several organisms including HCMV. The maintenance of the HCMV BAC in E. coli cells permits the rapid generation of recombinant viral genomes that can be used to produce viral progeny in cell cultures for the study of gene function. We optimized the Lambda-Red Recombination system to construct HCMV gene deletion mutants rapidly in the complete set of tested genes. This method constitutes a useful tool that allows for the quick generation of a high number of gene deletion mutants, allowing for the analysis of the whole genome to improve our understanding of HCMV gene function. This may also facilitate the development of novel vaccines and therapeutics. View Full-Text
Keywords: cytomegalovirus; BAC; Lambda-RED system; recombination; gene-deletion mutants cytomegalovirus; BAC; Lambda-RED system; recombination; gene-deletion mutants
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MDPI and ACS Style

García-Ríos, E.; Gata-de-Benito, J.; López-Siles, M.; McConnell, M.J.; Pérez-Romero, P. Optimization of a Lambda-RED Recombination Method for Rapid Gene Deletion in Human Cytomegalovirus. Int. J. Mol. Sci. 2021, 22, 10558. https://doi.org/10.3390/ijms221910558

AMA Style

García-Ríos E, Gata-de-Benito J, López-Siles M, McConnell MJ, Pérez-Romero P. Optimization of a Lambda-RED Recombination Method for Rapid Gene Deletion in Human Cytomegalovirus. International Journal of Molecular Sciences. 2021; 22(19):10558. https://doi.org/10.3390/ijms221910558

Chicago/Turabian Style

García-Ríos, Estéfani, Julia Gata-de-Benito, Mireia López-Siles, Michael J. McConnell, and Pilar Pérez-Romero. 2021. "Optimization of a Lambda-RED Recombination Method for Rapid Gene Deletion in Human Cytomegalovirus" International Journal of Molecular Sciences 22, no. 19: 10558. https://doi.org/10.3390/ijms221910558

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