Plasmid transformation is a foundational technique in life science research in fields such as molecular biology and protein science. Transformation is a process of horizontal gene transfer of exogenous DNA into a host cell (typically bacterium) where it can be cloned by the host cell in large numbers. While this technique can occur naturally, not all bacteria are capable of taking up exogenous DNA or do it at a sufficient rate to be tractable in the laboratory. Using chemicals or electrical pulses, however, one can generate artificially competent bacterial cells capable of uptaking exogenous DNA at high levels.
In this booklet, we provide a series of studies, protocols and reviews focusing on the theory, technique and applications of plasmid transformation. Through the presented article summaries and application note, we hope to educate researchers on important considerations and strategies to enhance the efficiency for successful plasmid DNA transformation.
What you will learn:
- The process of plasmid transformation and the key components required
- Considerations for plasmid transformation in E. coli
- Applications of plasmid transformation in other organisms through case studies
Contents:
- Overview of protein expression vectors for E. coli – Riggs et al. (2018)
- Generation of transformation competent E. coli – Renzette et al. (2011)
- Development of an in vivo methylation system for transformation of Ruminiclostridium cellulolyticum – Cheng et al. (2022)
- Introduction of plasmid DNA into cells – Seidman et al. (2001)
- Harnessing the type I CRISPR-Cas systems for genome editing in prokaryotes – Xu et al. (2020)
- Plasmid DNA transformation in E. coli: considerations for essential lab equipment – Haupt and Wiggenhauser (2022)