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Updated: Nov 19, 2025

Genome-wide Gene Deletions in Streptococcus sanguinis by High Throughput PCR
Published on: November 23, 2012
Genomic deletions in Rhodococcus based on transformation of linear heterologous DNA
Divya Singhi1, Shabnam Parwin1, Preeti Srivastava1
1Department of Biochemical Engineering and Biotechnology, Indian Institute of Technology, New Delhi 110016, India.
Abstract:
Several genome engineering methods have been developed for Rhodococcus. However, they suffer from limitations such as extensive cloning, multiple steps, successful expression of heterologous genes via plasmid etc. Here, we report a rapid method for performing genomic deletions/disruptions in Rhodococcus spp. using heterologous linear DNA. The method is cost effective and less labour intensive. The applicability of the method was demonstrated by successful disruption of rodA and orphan parA. None of the disrupted genes were found to be essential for the viability of the cell. Disruption of orphan parA and rodA resulted in elongated cells and short rods, respectively. This is the first report demonstrating disruption of rodA and orphan parA genes by electroporation of heterologous linear DNA in Rhodococcus spp.
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