Content area
Abstract
Objective
To improve the transformation efficiency of Corynebacterium glutamicum cells with heterogenous plasmid DNA and single-strand DNA (ssDNA) using a methodology based on electro-transformation.
Results
A semicomplex hypertonic medium was selected with addition of glycine and dl-threonine to weaken cell walls and addition of Tween 80 and isonicotinic acid hydrazide to increase cytoplasmic membrane fluidity. Their contents were optimized by response surface methodology. Cell growth, electro-transformation buffer, and transformation protocol were also optimized. Temporary heating inactivation of the host restriction enzyme showed a significant effect. Finally, a high transformation efficiency of 3.57 ± 0.13 × 10^sup 7^ cfu/[mu]g DNA of plasmid and 1.05 × 10^sup 6^ Str^sup R^ cfu per 10^sup 9^ viable cells with a ssDNA was achieved.
Conclusion
The results shed light on the application in functional genomics and genome editing of C. glutamicum.