National Center for Microbiology, Instituto de Salud Carlos III Majadahonda, 28220 Madrid, Spain.
Int J Mol Sci. 2021 Sep 29;22(19):10558. doi: 10.3390/ijms221910558.
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.
人巨细胞病毒（HCMV）仍然是移植患者和新生儿发病的主要原因。然而，HCMV 基因组中编码的 282 多个基因的许多功能仍然未知。细菌人工染色体（BAC）技术的发展有助于包括 HCMV 在内的几种生物体的遗传操作。HCMV BAC 在大肠杆菌细胞中的维持允许快速生成重组病毒基因组，可用于在细胞培养物中产生病毒后代，以研究基因功能。我们优化了 Lambda-Red 重组系统，以在整套测试基因中快速构建 HCMV 基因缺失突变体。该方法构成了一种有用的工具，可快速生成大量基因缺失突变体，从而对整个基因组进行分析，以提高我们对 HCMV 基因功能的理解。这也可能有助于新型疫苗和疗法的开发。
